AN INSEMINATION DEVICE AND USES THEREOF

Field of the Invention

The present invention relates to gynecological instruments for artificial insemination. Specifically, it is set within artificial insemination devices that can be used by the users themselves.

State of the Art

Assisted insemination processes have become a common medical practice to help couples or single mothers to conceive and have children.

Nowadays, these kinds of practices are conducted using complex processes that require different types of equipment and stages to be executed. Generally, they are performed in clinics on a specific date, requiring coordination and transfers that, added to the natural anxiety about the treatment, cause stressful situations that may affect the ability of the person to be inseminated to conceive at that specific moment.

The assisted insemination process has several stages, including collecting the semen sample, selecting those spermatozoa with the best motility, morphological and genetic characteristics, and preparing the person to be inseminated for the insemination process itself. The sperm selection process, according to the techniques used today, requires the use of complex equipment in specialized centers which perform assisted reproduction treatments.

In the case of sperm selection devices for humans, various devices can be found in the state of the art. For example, the document WO2018/154169 describes a technology developed by the same inventors, consisting of two reservoirs separated by a sieved plate with channels that has a conical or similar shape. In said invention, the sample to be separated is deposited in the first reservoir, while in the second reservoir, a culture medium is added up to a level mark that ensures that a current is generated towards the first reservoir. In this device, the spermatozoa are separated, taking advantage of their mobility due to the phenomenon of rheotaxis and other mechanisms.

Another example, described in document US 8,535,622 B2, consists of a spermatozoa separation system based on their natural movement. Said invention comprises two chambers; the first is where the semen sample is deposited, which is delimited by a crenelated ring, and the second is where the medium that allows separation is added. Once the semen sample is placed in the first chamber, the medium that allows separation is added in the second chamber until it floods the first chamber and the separation of mobile cells occurs naturally. Likewise, document US 2019/0383722 discloses a system and method for sperm sorting. Said system comprises a housing and a microfluidic system supported by the housing. The system also features an inlet to the microfluidic system to add sperm and an outlet to the microfluidic system to collect sorted sperm. The microfluidic system provides a flow path for sperm from inlet to outlet and includes at least one channel that extends from inlet to outlet to allow the sperm, delivered to the microfluidic system, to progress from the inlet along the flow path to outlet. Said microfluidic system also comprises a filter that includes a plurality of micropores arranged in the flow path between the inlet and the outlet to cause spermatozoa to travel along the flow path and pass through the filter and against gravity to reach the outlet. All these systems allow a sperm selection that can be simply and easily executed with the advantage of not requiring complex instrumentation or trained operators.

On the other hand, various devices have been developed to facilitate assisted insemination. Such is the case of Ultimate Flex IUI Kit with Vaginal Camera Scope Speculum with Light; as well as other devices such as STORK OTC or those described in patent applications MX2019006283 of IN-NOMED ONE LLC (US) wherein a medical device for artificial insemination is provided, where the system is inserted into the cervical canal of the person to be inseminated and allows a cannula, which contains the sample to be inseminated, to pass through it. Said device also contains a series of elastomeric barriers that prevent the backflow of the inseminated sample. The device can be used by a medical team or by the person to be inseminated at home. The document CN201920696 ofTIANYU HAO; KUAN HAO, introduces a vaginal speculum that includes a color camera and a light. This device allows observation of the female vaginal area. In addition, it is worth mentioning the utility model CN201743665, which claims a cone-shaped system for visualizing the female genital area. Likewise, document W02014025404 from THE STORK IB2C INC (formerly RINOVUM WOMEN'S HEALTH, INC), discloses in its family of patents a system to promote fertility and a method of using it.

However, there is still a need for a device that allows executing both steps (sperm sorting and insemination) in a combined, safe and efficient way that is also suitable for use in a domestic environment and by untrained operators.

Brief Description of the Figures

Figure 1 is a representation of a drive mechanism in its initial position, in a preferred embodiment of the present invention

Figure 2 is a representation of said drive mechanism wherein an inner plunger has been actuated to pressurize a selected sperm reservoir. Figure 3 is a representation of said drive mechanism wherein an outer plunger has been actuated to close a semen injection port and a culture medium injection port.

Figure 4 is a representation of said drive mechanism wherein a cannula plug has been removed.

Figure 5 is a representation of said drive mechanism wherein an outer plunger has been actuated to deploy a discharge cannula out of a sterile cap.

Figure 6 is a representation of said drive mechanism wherein an inner plunger has been actuated to empty the content of a selected sperm reservoir and discharge it outside a discharge cannula.

Figure 7 is a perspective view of the control platform and disposable cartridge of the insemination device 1 described in Example 1, in accordance with the present invention.

Figure 8 is a perspective view of the control platform of the device described in Example 1.

Figure 9 is a side view of the insemination device 1 described in Example 1 with an adaptable rotating handle.

Figure 10 is a top view of the drive mechanism of the device described in Example 1.

Figure 11 is a longitudinal section of the drive mechanism of the device described in Example 1.

Figure 12 is a longitudinal section of the disposable cartridge of the device described in Example 1.

Figure 13 is a perspective view of the control platform, disposable cartridge and incubation base of the device described in Example 1.

Figure 14 is a longitudinal section of the disposable cartridge and drive mechanism which shows the selected sperm reservoir covered by the cover sleeve.

Figure 15 is a longitudinal section of the disposable cartridge and drive mechanism which shows the preloading of the retractable discharge cannula.

Figure 16 is a longitudinal section of the disposable cartridge and drive mechanism which shows the discharge cannula out of the sterile cap and the emptying of the selected sperm reservoir. Figure 17 is a longitudinal sectional of the disposable cartridge and drive mechanism which shows the actuation of the secondary injection plunger and emptying of the retractable discharge cannula.

Figure 18 is a perspective view of the control platform and disposable cartridge of the insemination device 2 described in Example 3, in accordance with the present invention,

Figure 19 is a perspective view of the control platform of the device described in Example 3.

Figure 20 is a longitudinal section of the control platform of the device described in Example 3. Figure 21 is a perspective view of the disposable cartridge of the device described in

Example 3.

Figure 22 is an exploded view of the disposable cartridge of the device described in Example 3.

Figure 23 is a longitudinal section of the disposable cartridge of the device described in Example 3.

Figure 24 is an exploded view of the drive mechanism of the device described in Example 3.

Figure 25 is a perspective view of the control platform, disposable cartridge and incubation base of the device described in Example 3. Figure 26 is a top, side and bottom perspective view of the control platform and disposable cartridge of the insemination device 2 described in Example 3.

Figure 27 is a longitudinal section of the disposable cartridge of the device described in Example 3. Detail A shows the position lock. Detail B shows the open semen and culture medium injection ports. Figure 28 is a perspective view of the inner plunger and outer plunger cap of the device described in Example 3 showing the mark on the outer plunger cap aligned with the 2-points mark on the inner plunger.

Figure 29 is a longitudinal section of the disposable cartridge of the device described in Example 3 wherein the semen and culture medium injection ports are closed. Figure 30 is a perspective view of the control platform and disposable cartridge of the insemination device 2 of Example 3 with a fully open speculum lid.

Figure 31 is a perspective view of the inner plunger and outer plunger cap of the device described in Example 3 showing the mark on the outer plunger cap aligned with the single- line mark on the inner plunger.

Figure 32 is a longitudinal section of the control platform and the disposable cartridge of the device described in Example 3 showing the inner and outer plungers in their final position.

Figure 33 is a boxplot showing the results of the functionality validation tests of the insemination device of the present invention.

Brief Description of the Invention

The insemination device, main objective of the present invention, comprises an inseminator; a sperm sorting chamber; and a drive mechanism which drives both the sperm sorting chamber and the inseminator; all integrated into the same device, Wherein the sperm sorting chamber comprises a semen injection port, a semen reservoir, a selection membrane, a selected sperm reservoir, and a culture medium injection port. And said selection membrane comprises at least one channel linking said semen reservoir with said selected sperm reservoir, preferably said selection membrane comprises a multiplicity of channels linking said semen reservoir with said selected sperm reservoir. Wherein said inseminator comprises a sterile cap and a discharge cannula, preferably retractable. And wherein said inseminator further comprises a camera, illumination means, a speculum lid and a speculum lid drive mechanism. Wherein said drive mechanism comprises compressing means for actuating the sperm selection chamber and discharging culture medium with selected spermatozoa, preferably at least one plunger, more preferably at least two telescopic plungers. Wherein said plunger slides inside the selected sperm reservoir.

Wherein said sperm sorting chamber, in a preferred embodiment, further comprises a separating membrane.

In an alternative embodiment of the present invention, said sperm sorting chamber further comprises a culture medium reservoir and said sperm sorting chamber further comprises an injection ports accessory. An embodiment of the present invention further comprises an electrical resistance; a temperature sensor; a temperature controller; the electronic components necessary to establish a wireless connection (WiFi or Bluetooth type) and run an "ad hoc" software application. Also could comprise a battery; a connector; cannula plug.

In one embodiment, the insemination device of the present invention comprises a disposable cartridge. Wherein said disposable cartridge comprises: a sperm sorting chamber, including a semen injection port, a semen reservoir, a selection membrane, a selected sperm reservoir, and a culture medium injection port; a separating membrane; a culture medium reservoir; a speculum lid and a speculum lid drive mechanism; at least a plunger (that could be at least two telescopic plungers); a sterile cap and a discharge cannula; a cannula plug.

A preferred embodiment of the insemination device of the present invention further comprises a control platform. Wherein said control platform comprises elements selected from the group consisting of a camera, illumination means, the electronic components necessary to establish a wireless connection (WiFi or Bluetooth type) and run an "ad hoc" software application, a handle, mechanical attachment to attach the disposable cartridge to the control platform, an electrical resistance, a temperature sensor, a temperature controller, a battery, a connector and its combinations.

A preferred embodiment of insemination device of the present invention further comprises an incubation base. Wherein said incubation base comprises elements selected from the group consisting of a connector; an electrical resistance and a temperature controller.

Another object of the present invention is a method of using the insemination device or an insemination method using the insemination device of the invention wherein it comprises, at least, the following steps: a. introducing the culture medium into the selected sperm reservoir and the semen sample into the semen reservoir, b. sealing the sperm sorting chamber to maintain watertightness, c. putting the device at rest at a temperature between 35 °C and 38 sc for at least 15 minutes, d. actuating the drive mechanism to empty the selected sperm reservoir containing the culture medium with selected spermatozoa, e. and subsequent discharge of said culture medium with selected spermatozoa outside the discharge cannula,

Detailed Description of the Invention

The insemination device, the main object of the present invention, comprises an inseminator; a sperm sorting chamber; and a drive mechanism which drives both the sperm sorting chamber and the inseminator; all integrated into the same device.

The term "inseminator" refers to the parts of the device that are involved in detecting the cervix and discharging selected spermatozoa into the uterus.

Said inseminator comprises at least: a sterile cap, which is introduced into the vaginal canal; and a discharge cannula that can be inserted into the uterus to discharge the selected spermatozoa.

In one embodiment of the invention, said discharge cannula is retractable, so it is hidden inside the sterile cap and can be deployed by actuating the drive mechanism at the moment of performing the insemination.

The term "sperm sorting chamber” refers to the parts of the device that are involved in selecting motile cells, in particular spermatozoa, from a semen sample.

Said sperm sorting chamber comprises at least a semen injection port, a semen reservoir, a selection membrane, a selected sperm reservoir, and a culture medium injection port.

Said sperm sorting chamber forms a space wherein spermatozoa are selected from a semen sample and comprises a semen injection port that allows the user to incorporate a semen sample into said semen reservoir. Said semen reservoir is linked to a selected sperm reservoir by a selection membrane containing at least one communication channel between both reservoirs. Preferably, the selection membrane contains a multiplicity of channels through which spermatozoa can swim. More preferably, the selection membrane has channels whose inlet openings have greater diameters than the middle section of said channels, In a preferred embodiment of the invention, said selection membrane is that described in patent document W02018/154169, which is incorporated herein by reference.

In turn, said culture medium injection port allows the user to incorporate culture medium into said selected sperm reservoir. The sperm sorting chamberfurther comprises a separating membrane. Said separating membrane breaks when a certain amount of pressure is applied on it, allowing the transfer of culture media with selected spermatozoa from the selected sperm reservoir to the discharge cannula at the desired moment.

The term "drive mechanism" refers to the parts of the device that are involved in the operation of both the sperm sorting chamber and the inseminator. In other words, it refers to a mechanism that, when actuated, allows the following actions to be performed: actuate the sperm sorting chamber, meaning at least, sealing the sperm sorting chamber to maintain watertightness within said sperm sorting chamber, and emptying the selected sperm reservoir containing the culture medium with selected spermatozoa; and actuate the inseminator, meaning at least, discharging said culture medium with said selected spermatozoa outside through the cannula.

The operation of said drive mechanism allows untrained users to conduct the sperm selection process and the discharge of selected spermatozoa out of the device in a simple way and in a domestic environment. Said drive mechanism comprises compressing means for actuating the sperm selection chamber and discharging culture medium with selected spermatozoa,

Preferably, in one embodiment of present invention, said compressing means comprise at least one plunger that slides into the selected sperm reservoir, allowing emptying the selected sperm reservoir containing the culture medium with selected spermatozoa, and subsequent discharge of said culture medium with selected spermatozoa outside the discharge cannula. In a more preferred embodiment, said compressing means comprise at least two telescopic plungers.

In a preferred embodiment of the invention, the selection membrane has a cylindrical geometry wherein the plunger or piston inserted into the selected sperm reservoir have a circular section. In other embodiments of the invention, said selected sperm reservoir and the respective plunger and/or piston inserted into it have matching sections selected from the group comprising, but not limited to, parallelograms, hexagons, trapezoids and ellipses.

In turn, the insemination device of the present invention can comprise a series of accessories that contribute to a simple operation of the device and facilitate the insemination procedure.

In one embodiment of the invention, said inseminator further comprises a camera that allows filming the insemination procedure and locating the cervix. In another embodiment of the invention, said inseminator further comprises illumination means, to illuminate the vaginal canal. In a preferred embodiment, said illumination means comprise at least one LED light.

In another embodiment, the inseminator comprises a light intensity control that adapts accordingly to the user's needs.

In another embodiment, the inseminator further comprises the electronic components necessary to establish a wireless connection (WiFi or Bluetooth type) and run an "ad hoc" software application. According to this embodiment, the device can establish a wireless connection (WiFi or Bluetooth type) and run an ^" ad hoc ^" software application that, through image recognition, guides and assists the user to identify the cervix and perform an intrauterine insemination procedure. The user guidance and assistance process can be conducted by an application installed on a smartphone/tablet/laptop chosen by the user, doctor, assistant, or person in charge of operatingthe device, in accordance with the different telemedicine assistance systems currently known, including those performed via streaming.

In another embodiment, the ad hoc developed application is able to photograph and record the performed procedure, store the information to be analyzed later and send it where it is required. In a more preferred embodiment, it comprises a WiFi transmitter module capable of sending the displayed images anywhere in the world. The ability to transmit these images in real-time enables any professional anywhere in the world to assist in the procedure to anyone who is performing it. Likewise, the versatility of the device allows it to be used for cervical cancer screening and diagnosis, remote Pap smear testing, or any other procedure that requires the application of a cytobrush.

In another embodiment, the distal end of the inseminator is able to rotate on the vertical plane by an angle up to ± 90 ° to facilitate the entry of the discharge cannula into the cervix located in an anteverted or retroverted uterus.

In another embodiment, the inseminator further comprises a mechanism that directs the discharge cannula by modifying its trajectory on the vertical plane at an angle up to ± 90 ° to facilitate the entry of the discharge cannula into the cervix located in an anteverted or retroverted uterus.

In another embodiment of the invention, said inseminator further comprises a speculum lid and a speculum ltd drive mechanism. Said speculum lid emulates the operation of a speculum and gives the device the ability to separate the vagina! cavity walls, facilitating the identification of the cervix and consequently, the insemination procedure. Said drive mechanism is similar to those typically used in commercially available disposable specula. The speculum lid drive mechanism comprises: a screw; a perforated tab with a threaded hole; and a grooved tab.

In another embodiment, said inseminator device further comprises a handle. In a preferred embodiment, said handle can be rotated in order to adapt the grip to the position that best suits the users needs and facilitates its storage.

In an embodiment of the invention, said sperm sorting chamber further comprises a culture medium reservoir. Said culture medium reservoir consists of a chamber wherein said culture medium is introduced before being transferred to the selected sperm reservoir.

The sperm sorting chamber may contain the selection medium already integrated into it. Therefore, in this case, at least the following possibilities can be envisaged:

Culture medium outside the sperm sorting chamber: it is introduced into the culture medium reservoir or selected sperm reservoir before performing the selection. ii. Lyophilized culture medium, integrated into a solid pellet within the culture medium reservoir or selected sperm reservoir: when the device is used, the appropriate fluid (WFI water, distilled water, aqueous solution, to be defined) is incorporated, dissolving the pellet and allowing to proceed with the selection. iii. Selection medium integrated in a separate cartridge, in liquid format: at the moment of use, said cartridge is placed in the corresponding space, and a security barrier is broken to proceed with the selection. iv. Selection medium integrated in a separate cartridge, in solid format: at the moment of use, the medium pellet is dissolved with the appropriate fluid (WFI water, distilled water, aqueous solution, to be defined). Then, said cartridge is placed in the corresponding space, and a security barrier is broken to proceed with the selection.

The terms "culture medium" and "selection medium" refer to the fluid to which motile spermatozoa access after leaving the seminal plasma by themselves.

In another embodiment, the sperm sorting chamber further comprises a temperature sensor located near the sperm sorting chamber to measure its temperature. In a preferred embodiment, said temperature sensor comprises a thermocouple.

In another embodiment, the sperm sorting chamber further comprises heating means and a temperature controller to provide heat and maintain a stable temperature within the semen reservoir and the selected sperm reservoir. In a preferred embodiment, said heating means comprise an electrical resistance powered by a battery or by an external power source. More preferably, said heating means comprise an electrical resistance which is powered externally, in this way it does not rely on batteries to provide heat and excessive battery draining due to the high consumption of electrical resistances is avoided.

In yet another embodiment of the present invention, the sperm sorting chamber comprises an injection ports accessory that physically limits the depth to which syringes and/or pipettes used to introduce fluids (i.e., semen, culture medium, WFI water, etc.) can be inserted. In this way, possible damage to the membrane or other inner structures of the sperm sorting chamber Is avoided. In a preferred embodiment, said injection ports accessory is disposable and consists in a solid piece with through holes which is placed in such a way that the holes of the injection ports accessory align with the semen and culture medium Injection ports. In a preferred embodiment, a Pasteur pipette is used to incorporate around 3 ml of semen sample, and a 1 ml tuberculin syringe is used to incorporate around 0.85 ml of culture medium.

In another embodiment, said insemination device further comprises at least one battery for powering the electronic components that require it.

In another embodiment, said insemination device further comprises connectors for establishing a connection to the domestic electrical network in order to be able to power the electronic components that require it and/or recharge its batteries.

In another embodiment of the present invention, the insemination device further comprises a cannula plug to prevent accidental rupture of the separating membrane when pressure is applied to the selected sperm reservoir.

Figures 1 to 6 show different steps of a drive mechanism of one embodiment of the present invention, actuating a sperm sorting chamber and an inseminator. In this sense, Figure 1 depicts an initial position wherein a semen injection port and a culture medium injection port are open to allow the incorporation of a semen sample and culture medium. In Figure 2 an inner plunger has moved a small distance, pressurizing a selected sperm reservoir and consequently flooding the channels of a selection membrane with culture medium. In Figure 3 an outer plunger has advanced a small distance and both the semen injection port and the culture medium injection port have been closed to seal the sperm sorting chamber and maintain water tightness. In Figure 4 a cannula plug has been removed, which has to be done before the device is inserted into a vaginal canal. In Figure 5 an outer plunger has been moved and a discharge cannula has been deployed out of a sterile cap. In Figure 6 an outer plunger has advanced, emptying the content of a selected sperm reservoir, and discharging it outside a discharge cannula. Another object of the present invention, is a sperm selection and insemination method that comprises, at least, the following steps: a. introducing the culture medium into the selected sperm reservoir and the semen sample into the semen reservoir, b. sealing the sperm sorting chamber to maintain water tightness, c. putting the device at rest at a temperature between 35 °C and 38 °C for at least 15 minutes, d. actuating the drive mechanism to empty the selected sperm reservoir containing the culture medium with selected spermatozoa, e. and subsequent discharge of said culture medium with selected spermatozoa outside the discharge cannula.

In a preferred embodiment, the step at a temperature of about 35 and 38 °C lasts for at least 30 minutes. In a more preferred embodiment, the step of putting the device to rest is conducted at a temperature between 36 °C and 37 °C. In an even more preferred embodiment, the step of putting the device to rest is conducted at 37 “C.

Application examples

Example 1

Insemination device 1

An embodiment of the present invention is described in detail below.

In order to materialize the present invention, one embodiment of the invention comprises a control platform (1), a disposable cartridge (2), and an incubation base (4) that interact with each other in order to execute the actions needed to perform the insemination procedure.

Control platform (1)

The control platform is a reusable support that can be attached to the disposable cartridge, as illustrated in Figure 7, and allows the user to manipulate the insemination device. The control platform comprises a camera; illumination means; the electronic components necessary to establish a wireless connection (WiFi or Bluetooth type) and run an "ad hoc" software application; a handle; mechanical attachment mechanism, to attach the disposable cartridge to the control platform; a temperature sensor; a battery; and a connector.

In more detail, said control platform comprises:

1. A camera (101), which is located at the front end of the device, according to what is illustrated in Figure 8.

2. Illumination means comprising an LED light (102), which is located at the front end of the device.

3. The electronic components necessary to establish a wireless connection (WiFi or Bluetooth type) and run an "ad hoc" software application.

4. A handle (103) that can be rotated to adapt the grip to the position that best suits the users needs and facilitates its storage, as illustrated In Figure 9.

5. A mechanical attachment mechanism, to attach the disposable cartridge to the control platform. Said mechanical attachment mechanism comprises two clamping arms (104).

6. A battery for powering electronic components that require it, which is located inside the handle.

7. A magnetic connector (105) that establishes an electric connection with the incubation base platform.

8. A temperature sensor comprising a thermocouple which is located near the sperm sorting chamber when the disposable cartridge is attached to the control platform.

In addition, the insemination device comprises a drive mechanism (3) that drives the inseminator and the sperm sorting chamber through plungers inside the disposable cartridge. In this embodiment of the invention, the control platform comprises three telescopic plungers; which, according to their functions, are called:

(301) Drive plunger for membrane sealing and sperm sorting chamber push.

(302) Primary injection drive plunger.

(303) Secondary injection drive plunger.

As illustrated in Figures 10 and 11, in this embodiment of the invention, said mechanism comprises a selector ring (304) which has a safety pin (305) that is positioned within the guiding grooves (306) of the telescopic plungers (301, 302 and 303). The safety pin (305) that works jointly with the selector ring (304) can only rotate together. In this way, the action ofturningthe ring (304) defines which guide (306) will be able to slide and consequently which of the plungers (301, 302, and/or 303) will be able to advance.

The three plungers (301, 302, and 303) have guides along their axes with paths that allow the selector ring (304) to rotate in pre-defined degrees. These guiding grooves (306) can overlap depending on the need to actuate the device, enabling the movement of more than one plunger at the same time. Therefore, each time the selector ring is rotated, it chooses which guide, and therefore which section of the plunger will be able to move a predetermined distance. In other words, said plungers are part of a drive mechanism, which depending on the position selected by the ring (304), can move together or separately.

According to the present embodiment of the invention, the insemination device comprises further a sliding button (307), which in one position functions as a lock for the selector ring (304) and in the other position interacts with a component inside the disposable cartridge to close or seal the semen and the culture medium injection ports.

Disposable cartridge (2)

Said disposable cartridge is the part of the device that is inserted into the vaginal canal and, according to the present embodiment, comprises the sperm sorting chamber and the inseminator.

In more detail, said disposable cartridge, illustrated in Figure 12, comprises;

1. A sterile cap (201). Said cap comprises a series of openings: a. Two openings on the top to allow the addition of a semen sample and culture medium to the sperm sorting chamber through the semen injection port (202) and the culture medium injection port (203), respectively.

Likewise, at the distal end of the device, it comprises a series of openings: b. An opening that allows the exit of a retractable discharge cannula (204) from the interior of the sterile cap (201) for its insertion into the uterine cavity. c. An openingthat allows the camera (101) to visualize the vaginal canal. d. An opening that allows the LED (102) to illuminate the vaginal canal adequately,

2. A culture medium reservoir (205). Said reservoir comprises a culture medium injection port (203) which is open before the culture medium is injected, and closes afterwards. 3. A semen reservoir (206) comprising a concentric cylindrical selection membrane (207) that allows the selection of spermatozoa suitable for insemination. Said semen reservoir (206) comprises a semen injection port (202) which is open before the semen sample is injected, and closes afterwards. Said selection membrane (207) comprises a multiplicity of channels that allow the self-selection of those spermatozoa with adequate characteristics. In this embodiment of the present invention, said selection membrane is that described in patent document WO2018/154169,

4. A selected sperm reservoir (208), which is separated from the semen reservoir (206) by the selection membrane (207).

5. A sliding gate (209), actuated by (307), that closes the semen and culture medium injection ports (202 and 203) to maintain water tightness within the sperm sorting chamber and prevent the fluids from escaping from said sperm sorting chamber through said ports by the actuation of the plungers.

6. A retractable discharge cannula (204), next to the selected sperm reservoir (208), which communicates to said reservoir (208) through an opening.

7. A separating membrane (210) that separates the selected sperm reservoir (208) from the cannula (204). Said separating membrane breaks when a certain amount of pressure is applied on it, allowing the transfer of culture media with selected spermatozoa from the selected sperm reservoir to the cannula at the desired moment.

8. An internal plunger for membrane sealing and sperm selection chamber push (308) that is actuated by (301).

9. A cover sleeve (309) that, by actuation of (301), covers the selected sperm reservoir (208). in this way, the loss of selected sperm through the selection membrane is avoided when pressure is generated within the selected sperm reservoir (208) during the filling of the retractable discharge cannula (204).

10. An internal primary injection plunger (310), actuated by (302), which pushes the piston (311).

11. An internal secondary injection plunger (312), actuated by (303), which is inserted into the discharge cannula to discharge it completely.

Incubation base (4)

The incubation base, illustrated in Figure 13, comprises a magnetic connector (401) to supply electrical power to the batteries, an electrical resistance to provide heat to the sperm sorting chamber during the selection procedure, and a temperature controller to keep the temperature within the desired range. In turn, the attached disposable cartridge and control platform can rest on said incubation base. In this vein, the incubation base provides physical support and allows the device to remain still during the sperm selection process.

Example 2

A method of use of the embodiment described in Example 1 is exemplified below.

The insemination device is operated as indicated in the following steps:

0. Install the app on a smartphone, tablet, or laptop. Turn on the device and connect it to the app. By starting the insemination cycle in the app, it will guide the userthrough the entire procedure.

1. Lift the safety pin (305) to release the telescopic plunger mechanism and extend plungers (301, 302, and 303). Set the selector ring (304) to position "I". Return the pin (305) to its original position.

2. Attach the disposable cartridge (2) to the control platform (1) sliding it until a click is heard as a consequence of the mechanical attachment between the clamping arms (104) and the disposable cartridge (2).

3. Add culture medium through port (203) and add semen sample through port

(202).

4. Close the injection ports (202 and 203) by actuating the sliding button (307).

5. Place the device in the incubation base (4), where the sample is heated in a controlled and automatic manner for at least 15 minutes to favor the sperm selection process.

6. Once the selection process is completed, the insemination process can proceed. The process is guided by the application installed on the smartphone, tablet or laptop.

7. The selector ring (304) is rotated to position "II", and the plunger (301) is actuated to cover the selected sperm reservoir with the cover sleeve (309), which is pushed by the movement of the plunger, as illustrated in Figure 14. The selector ring is responsible for defining which plunger can be moved while limiting the travel of the other parts.

8. The selector ring (304) is turned to position "III", which enables movement of the primary injection drive plunger (302), limiting it to a short travel, This movement preloads the cannula (204) with the selected sperm while removing the air occupying the interior of the discharge cannula, preventing air from being injected before the sample (Figure 15).

9. At this point, the person to be inseminated must adopt a gynecological position. Remove the device from the incubation base, proceed to lubricate the exterior of the sterile cap (201), insert the device into the vaginal canal, and follow the instructions provided by the application to detect the cervix.

10. Once the cervix is detected, the selector ring (304) is placed in position "IV" to enable the subsequent movement of the plunger (301). Actuate (301) so that the retractable cannula (204) exits the sterile cap (201) and is inserted into the uterus.

11. Subsequently, the selector ring (304) is placed in the following position (position "V") to proceed with the insemination, and the primary injection drive plunger (302) is actuated, discharging the content of the selected sperm reservoir, as shown in Figure 16.

12. Finally, and to take advantage of the entire volume of the selected sperm, the selector ring is turned to its last position (position "VJ"), which releases the secondary injection drive plunger (303). As illustrated in Figure 17, the plunger (303) pushes the internal secondary injection plunger (312) into the discharge cannula, emptying it completely.

Example 3

Insemination device 2

Another embodiment of the present invention is described in detail below.

The present embodiment of the insemination device comprises a disposable cartridge, a control platform, and an incubation base that interact with each other in order to execute the actions needed to perform the insemination procedure.

Control platform (1)

The control platform is a reusable support that can be attached to the disposable cartridge, as illustrated in Figure 18, and allows the user to manipulate the insemination device. In a preferred embodiment the control platform comprises a camera, illumination means, the electronic components necessary to establish a wireless connection (WiFi or Bluetooth type) and run an "ad hoc" software application, a handle, mechanical attachment to attach the disposable cartridge to the control platform, an electrical resistance, a temperature sensor, a temperature controller, a battery, and a connector. In more detail, said control platform comprises:

1. A camera (101), which is located at the front end of the device, according to what is illustrated in Figure 19.

2. Illumination means comprising two LED lights (102), which are located at the front end of the device.

3. The electronic components necessary to establish a wireless connection (WiFi or Bluetooth type) and run an "ad hoc" software application.

4. A handle (103) that can be rotated to adapt the grip to the position that best suits the users needs and facilitates its storage.

5. A mechanical attachment mechanism, to attach the disposable cartridge to the control platform. Said mechanical attachment mechanism comprises one clamping arm (104),

6. A battery for powering electronic components that require it, which is located inside the handle.

7. A magnetic connector (105) that establishes an electric connection with the incubation base platform,

8. A sliding lock (106) that locks the handle in the desired position, as illustrated in Figure 20.

9. An electrical resistance (107) to provide heat to the sperm sorting chamber.

10. A temperature sensor and a temperature controller, to keep the temperature inside the sperm sorting chamber within the desired range. Said temperature sensor comprises a thermocouple which is located near the sperm sorting chamber when the disposable cartridge is attached to the control platform,

11. A perforated tab (108) with a hole that has a threaded inner surface and a screw (109) which are part of the speculum lid drive mechanism that opens and holds in position the speculum lid.

12. An on-off button (110) that turns on or off the electronic components of the device.

13. A release button (111) that allows detaching (2) from (1).

Disposable cartridge 12

Said disposable cartridge is the part of the device that is inserted into the vaginal canal and, according to the present embodiment, it comprises a sperm sorting chamber, an inseminator, and the drive mechanism (3).

In more detail, said disposable cartridge, illustrated in Figures 21 to 23, comprises: 1. A sterile cap (101). Said cap comprises a series of openings: a. Two openings on the top to allow the addition of semen and culture medium to the sperm selection cartridge through the semen injection port (202) and the culture medium injection port (203), respectively.

Likewise, at the distal end of the device, it comprises a series of openings: b. An opening that allows the exit of the retractable discharge cannula (204) from the interior of the sterile cap (201) for its insertion into the uterine cavity. c. An opening that allows the camera (101) to visualize the vaginal canal. d. Two openings that allow the LEDs (102) to illuminate the vaginal canal adequately.

2. A semen reservoir (206) comprising a concentric cylindrical selection membrane (207) that allows the selection of spermatozoa suitable for insemination. Said semen reservoir (206) comprises a semen injection port (202) which is open before the semen sample is injected, and closes afterwards. Said selection membrane (207) comprises a multiplicity of channels that allow the self-selection of those spermatozoa with adequate characteristics. In this embodiment of the present invention, said selection membrane is that described in patent document WO2018/154169.

3. A selected sperm reservoir (208), which is separated from the semen reservoir

(207) by the selection membrane (207). Said reservoir comprises a culture medium injection port (203) which is open before the culture medium is injected, and closes afterwards.

4. A retractable discharge cannula (204), next to the selected sperm reservoir

(208), which communicates to said reservoir (208) through an opening.

5. A separating membrane (210) that separates the selected sperm reservoir from the cannula (204). Said separating membrane breaks when a certain amount of pressure is applied on it, allowing the transfer of culture media with selected spermatozoa from the selected sperm reservoir to the discharge cannula at the desired moment.

6. A cannula plug (211) that prevents accidental rupture of the separating membrane when pressure is applied to the selected sperm reservoir.

7. A septum (212), located between the sterile cap and an outer plunger that seals the semen and culture medium injection ports. Said septum is pierced by the Pasteur pipette and the tuberculin syringe at the moment of adding semen sample and culture medium to the sperm sorting chamber. 8. A speculum lid (213). Said speculum lid is attached to the sterile cap through a hinge located at the end of the disposable cartridge closest to the handle (when the disposable cartridge is attached to the control platform).

9. A grooved tab (214), which is attached to said speculum lid and is part of the speculum lid drive mechanism. Said speculum lid drive mechanism imitates those typically used in disposable specula commercially available wherein said screw is placed in said perforated tab (108) with a threaded hole, the screw is then turned and pushes the grooved tab attached to the speculum lid, lifting said speculum lid as a result.

10. An injection ports accessory (215) that physically limits the depth to which the syringe and the pipette used to introduce fluids can be inserted. Said injection ports accessory is placed in a slot on top of the disposable cartridge. Said slot is placed in a way that the holes of the injection ports accessory align with the semen and culture medium injection ports.

Said disposable cartridge further comprises the drive mechanism (3) that allows the different actions to be performed within said disposable cartridge (2). Said drive mechanism, illustrated in Figure 24, comprises two telescopic plungers and a knob; which, are called:

(313) Inner plunger.

(314) Outer plunger.

(315) Command and selection knob.

According to this embodiment, said inner plunger (313) comprises a segment with a reduced section and a helicoid (316). Said helicoid (316) interacts with an outer plunger cap (317) causing that when the first counterclockwise turn of the command and selection knob (315) is executed, the inner plunger (313) advances a few millimeters, pressurizing the selected sperm reservoir and forcing the selection medium to flood the membrane channels.

Said inner plunger (313) further comprises visual and tactile indicators consisting of dots and lines which are engraved on its visible surface. Said indicators align with an embossed line located at the outer plunger cap (317), giving visual and tactile information about the different positions of the drive mechanism.

In addition, the inner plunger (313) has a guiding groove (306). When the command and selection knob is rotated counterclockwise for the second time, a tab located on the inner surface of the outer plunger cap (317) falls into the guiding groove and, in consequence, said inner plunger is able to advance a predetermined distance.

The outer plunger (314) further comprises indentations on its outer surface. Said indentations interact with a bump located on the inner surface of the sterile cap creating a position lock (318). Said position lock (318) gives feedback to the user and indicates where the drive mechanism has locked into a new position after pushing the command and the selection knob,

In turn, the whole drive mechanism is designed in such a way that, when operating the device according to the provided instructions, the user will feel and hear a click every time that the device locks in a new position.

Incubation base (4)

The incubation base, illustrated in Figure 25, comprises a magnetic connector (401), to supply electrical power to the batteries and to the electrical resistance. In turn, the attached disposable cartridge and the control platform can rest on said incubation base, in this vein, the incubation base provides physical support and allows the device to remain still during the sperm selection process.

In addition, the nose of the disposable cartridge has an ergonomic shape that facilitates the insertion of the device into the vaginal canal. In turn, said nose comprises a slit which functions as drainage of vaginal fluids, preventing them from accumulating in the cavity formed by the nose and obstructing the view of the camera (see Figure 26).

In the present document, terms "nose", "insemination device nose" or " disposable cartridge nose' ¹ are understood as the end of the device that is inserted into the vaginal canal.

Example 4

A method of use of the insemination device 2 described in Example 3 is exemplified below.

The initial position of the device is with the telescopic plungers (313 and 314) fully extended, this position is set by the position lock (318) that can be seen in detail A of Figure 27. In said initial position, the culture medium and semen injection ports (203 and 202) are open so that the fluids can be introduced into the sperm sorting chamber, as shown in detail B of Figure 27.

The device is operated as indicated in the following steps:

0. Install the app on a mobile device or laptop. With the device placed in its base (4), turn on the device and connect with the app. By starting the insemination cycle in the app, it will guide the user through the entire procedure.

1. Using a 1 ml tuberculin syringe, take 0.85 ml of selection medium from the vial, introduce the culture medium through the culture medium injection port (203), indicated with a drop icon.

2. Rotate the command and selection knob (315) counterclockwise to match the mark on the outer plunger cap (317) with the 2-points mark on the inner plunger (313), by doing this the user will feel that the plunger locks in position. This movement forces the inner plunger (313) to advance slightly and pressurize the selected sperm reservoir filled with the culture medium in step 1 (see Figure 28). At this point, the channels of the selection membrane will flood with the culture medium.

3. Using a Pasteur pipette, take 3 ml of the previously collected and homogenized semen sample and introduce it through the semen injection port (202), indicated with a sperm icon.

4. Push the command and selection knob (315) in order to move longitudinally the outer plunger (314) to the next point of the position lock indicated in detail A of Figure 27. In this position, the holes of the sterile cap (201) and outer plunger (314) that conform the semen and culture medium injection ports (202 and 203) become misaligned, closing said ports (see Figure 29).

5. At this point, the device must be left to rest for 30 minutes. During this time the incubation of the introduced sample and the sperm selection process will occur. The application will notify the user when the procedure has concluded.

6. After 30 minutes of incubation, take the device and remove the cannula plug (211).

7. The person to be inseminated must adopt a gynecological position. Lubricate the sterile cap (201) and the speculum lid (213) and insert the device into the vaginal canal, following the instructions provided by the app for detecting the cervix.

8. Once the device is inside the vaginal canal, open the speculum lid (213) by turning the screw (109) (see Figure 30).

9. Once the cervix has been detected, push the command and selection knob (315) so as to advance the outer plunger (314) until it stops and the cannula has been inserted into the cervix. 10. Rotate the command and selection knob (315) counterclockwise to the next position so that the mark on the outer plunger cap (317) is aligned with the single-line mark on the inner plunger (313). In this position the inner plunger is released and can advance longitudinally (see Figure 31).

11. Push the command and selection knob (315) so as to advance the inner plunger (313) until it stops. Atthis point both plungers (313 and 314) will have advanced to their final travel, as shown in Figure 32.

12. Retract the speculum lid (213) by unscrewing the screw (109). Gently remove the device and discard the sperm sorting chamber by pressing the release button (111) and pointing it towards a waste basket.

Example 5

Functionality validation tests of the insemination device of the present invention

Objective

The objective of the experiment was to evaluate the efficiency to separate spermatozoa from seminal plasma of the device of the present invention, and to evaluate the quality of recovered spermatozoa in terms of their morphology, motility and physiological parameters.

The separation efficiency of spermatozoa from the seminal plasma of the device of the present invention was compared with standard laboratory separation methods. In particular, the sperm recovery efficiency of the device of the present invention, the swim up technique and the density gradient separation technique, were compared. The percentage of cells recovered with each method {compared to the concentration in seminal plasma) and the percentage of motile cells was evaluated. Likewise, the morphology, kinetic parameters and physiological aspects were compared between the cells recovered by the previously mentioned methods (i.e., device of the present invention, swim up technique and density gradient separation technique).

Studied population. The study was performed using human semen samples considered as normozoospermic, teratozoospermic or oligozoospermic, according to WHO classification (WHO 2010), provided by men between 18 and 45 years of age. Samples with other sperm quality were not included in the study. Samples were collected between February and May 2021 Ethical considerations. The present study was conducted in accordance with the Helsinki Declaration. The human semen samples used during the experiments were collected from persons who have provided their written consent.

Sample collection. For the present study, the samples used presented a minimum sperm count of 20xl0 ⁶ sperm with at least a 40 % of progressive sperm.

Separation in the insemination device 2 of Example 3 (DEV). The sperm selection process was conducted following the steps given in Example 4. The selected sperm reservoir was filled with 0.85 ml of selection medium using a 1 ml tuberculin syringe and 3 ml of semen sample was introduced into the sperm reservoir. The device was left to incubate at rest on its incubation base for 30 min at 37 °C. Then, the sample with selected spermatozoa in the recovery medium was stored in an oven at 37 "C with 5% CO2 until testing.

Swim up separation (SWUP). The procedure was performed as described by WHO (WHO 2010). Briefly, 0.5 ml of semen was introduced into a sterile 15 ml conical tube and 1.2 ml of selection medium was added over it, gently and without mixing. The tube was incubated for 1 hour in a tilted position at 45 degrees in an oven at 37 °C with 5% CO2 in air. The tube was then gently placed upright and 1 ml of the supernatant medium was taken. This aliquot with motile cells was diluted with 8 volumes of modified HTF medium and centrifuged for 5 minutes at 500 xg. Finally, the supernatant was removed and the cells were resuspended in 0.5 ml of the same medium. The cells were kept in an oven at 37 °C with 5% CO2 until testing.

Density gradient separation (DGC). The procedure was performed as described by WHO (WHO 2010). Briefly, 1 ml of Percoll 80% was pipetted into a sterile 15 ml cone-based tube. Then, 1 ml of 40% Percoll was gently added on top of the 80% layer, taking care not to disturb the interface between the layers. Subsequently, 0.5 ml of semen was gently deposited on top of the gradient and the tube was centrifuged at 500 xg for 20 minutes. Then, 0.8 ml of the 80% fraction containing the motile cells were recovered and placed in a conical tube containing 8 ml of selection medium. The cells were then centrifuged at 500 xg for 5 minutes. The supernatant was removed and the cells were resuspended in 0.5 ml of HTFm medium. The cells were then kept in an oven at 37°C with 5% CO2 until testing.

Sperm concentration assessment. The sperm count was conducted according to WHO (WHO 2010) using a Neubauer hemocytometer. Briefly, an aliquot of the recovered sperm suspension was taken and fixed with 1% formalin. The Neubauer chamber was then loaded and the number of cells present in the central grid of the chamber was determined using a brlghtfield microscope at 40x (Olympus CX41). Cell concentration was then determined as follows: Concentration = cells number x dilution factor x 10000.

Sperm morphology assessment. Sperm morphology was evaluated by bright field microscopy according to the criteria defined by WHO (WHO 2010) in at least 200 spermatozoa, which had previously been subjected to Papanicolaou staining (WHO 2010).

Evaluation of sperm motility and kinetic parameters, spermatozoa were recorded at 100 frames per second using a digital camera (Andor Zyla 4.2) coupled to a phase contrast microscope (Nikon). From the images, the percentage of motile cells was determined and the trajectories of at least 150 spermatozoa per treatment were obtained using the Particle Tracker plugin of ImageJ software (NIH). From the trajectories, the linear and curvilinear velocity, as well as the movement pattern, were determined individually and by populations, using the paNoel vl.O software (UNC).

Evaluation of DNA integrity using the Haiosperm technique (Fernandez et al. 2005). A commercial Haiosperm G2 kit was used and the process was performed according to the manufacturer's instructions. Briefly, intact spermatozoa were embedded in an inert agarose microgel on a slide pretreated for this purpose. Then, an acid treatment was applied to denature the DNA. Next, a lysis treatment was performed to allow the DNA to unfold. The nucloids were then stained and the size of the halo of unfolded DNA was assessed by microscopy. Cells with intact DNA have large halos and those with damaged DNA have small halos or no DNA at all. The percentage of cells with intact DNA was determined by assessing the percentage of large halos in at least 200 nucleoids.

Determination of relative DNA content. The separation system is based on sperm motility, so it does not influence the distribution of the proportion of cells carrying X or Y chromosomes. The relative DNA content before and after separation was determined and compared with other separation methods to verify that there are no variations in the distribution of DNA content. For this purpose, spermatozoa were incubated with a solution containing 0.5 pg/ml Syber Green, for 30 min at 37 °C. The DNA content of the sperm population was evaluated by flow cytometry by analyzing at least 10000 events. The distribution data were analyzed using FlowJo software.

Statistical analysis. In order to determine the error distribution, at least 10 replicates of each experiment were performed and the data were analyzed with the Infostat software (UNC). For normal distributions, differences between treatments were determined by ANOVA and the post hoc DGC test, If the errors did not present a normal distribution, a nonparametric test was applied to determine the differences. Differences with a p<0,05 were considered significant. Results

Figure 33 exhibits the results of the functionality validation tests of the insemination device of the present invention. The graphs show the sperm count as well as the percentages of progressive motility, morphology and DNA fragmentation of the cells recovered by using the device of the present invention (DEV) in comparison with Swim up (SWUP) and density gradient (DGC) separation techniques, in addition, a neat semen sample (Neat) was analyzed; the obtained results are shown as a reference.

The results clearly show that the quality and quantity of the cells recovered using the insemination device of the present invention is similar or even better than those cells that were recovered by traditional techniques. In this vein, from Figure 33, it can be observed that the cells recovered using the device disclosed in this document (DEV) presented much less DNA fragmentation than DGC technique and similar to SWUP technique. In addition, the sperm count in the samples obtained using the device (DEV) were significantly higher than those obtained by the SWUP technique.

References Patent references

1. WO2018/154169; 2017-02-24; Selectivity, Repetto Constanza Julia, Repetto Flerberto Ernesto Elector, Selectivity Esease, Siletivi; Device and method for separating mobile cells.

2. US8535622; 2010-04-22; Lostu Bio Nymphaea, Lotus Bio; Sperm separation system.

3. US20190383722; 2014-11-19; Brigham & Womens Hospital; System and method for sperm sorting.

4. MX2019006283; 2016-11-30; Innomed One, Innomed One Liability, Innovative Medicine; Device and method for artificial insemination.

5. CN201920696; 2011-01-24; Hao Kuan, Hao Tianyu; Self-used vaginal speculum.

6. CN201743665; 2010-08-16; Hao Kuan, Hao Tianyu; Duckbill and probe combined vaginal speculum.

7. W02014/025404; 2013-03-15; Stork IB2C Group, Rinovum Womens Health, Stork IB2C, Stork IB2C Inc; Artifical insemination and vaginal insertion.

Non-patent references 1. WHO 2010. WHO Laboratory Manual for the Examination and Processing of Human Semen. Edited by Trevor G Cooper, John Aitken, Jacques Auger, HW Baker, Christopher L R Barratt, Hermann M Behre, Lars Bjorndahl, et al. Fifth Edit. Switzerland: WHO Press. http://whqiibdoc.who.int/publications/2Q10/9789241547789 eng.pdf?ua=l. 2. Fernandez, Jose Luis, Lourdes Muriel, Vicente Goyanes, Enrique Segrelles, Jaime

Gosalvez, Maria Enciso, Marie LaFromboise, and Christopher De Jonge. 2005. "Halosperm ^® Is an Easy, Available, and Cost-Effective Alternative for Determining Sperm DNA Fragmentation." Fertility and Sterility 84 (4): 860. https://doi.org/10.1016/i,fertnstert.2005.05.013.