Device for cryopreservation or thawing and recovery of biological tissues

Abstract

The present invention discloses a device for cryopreservation or thawing and recovery of biological tissues, which comprises: a biochip having a first microchannel for the flow of a solution, the first microchannel including a first solution inlet and a first solution outlet; a carrier including a second microchannel provided with a second solution inlet and a second solution outlet, and grid-like structures are respectively arranged at the upper and lower openings of the second solution outlet, and the grid-like structures confine the biological tissue in the second microchannel; wherein, the second solution inlet of the carrier is docked with the first solution outlet of the biochip. By means of the present invention, the delivery or removal of a cryoprotectant can be carried out in an orderly manner, effectively improving the efficiency of the cryopreservation or thawing and recovery operation of embryos / eggs.

Description

Technical Field

[0001] The present invention relates to the field of bioengineering technology, and particularly to a device for cryopreservation or thawing and recovery of biological tissues Background Art

[0002] Cell cryopreservation technology is an important technology in many biological fields. It is equally important to restore cells from a cryopreserved state to a normal metabolic state. The thawing and recovery process involves removing the cryoprotectant and replacing it with normal culture medium

[0003] The devices used in the prior art for embryo cryopreservation or thawing have problems such as complex operation and low efficiency Summary of the Invention

[0004] The main object of the present invention is to provide a device for cryopreservation or thawing and recovery of biological tissues, so as to solve the problems of complex operation and low efficiency existing in the devices of the prior art during embryo cryopreservation or thawing

[0005] The device for cryopreservation or thawing and recovery of biological tissues according to an embodiment of the present invention includes: a biochip having a first microchannel for solution flow, the first microchannel including a first solution inlet and a first solution outlet; a carrier including a second microchannel provided with a second solution inlet and a second solution outlet, and grid-like structures are respectively arranged at the upper and lower openings of the second solution outlet, and the grid-like structures confine the biological tissue in the second microchannel; wherein, the second solution inlet of the carrier is docked with the first solution outlet of the biochip

[0006] Wherein, the first solution inlet is arranged on the upper surface of the biochip, and the first solution outlet is arranged on the lower surface of the biochip

[0007] Wherein, the number of the first solution inlets is two

[0008] Wherein, the first microchannel is serpentine

[0009] Wherein, the grid-like structure includes a plurality of grid bars, and the distance between every two grid bars is less than the diameter of the biological tissue and greater than the diameter of solution molecules

[0010] Wherein, the carrier is closely attached to the lower side of the biochip, and the first solution outlet of the biochip covers the second solution inlet of the carrier

[0011] Wherein, the biochip does not cover the second solution outlet of the carrier

[0012] According to the technical solution of the present invention, during cryopreservation or thawing and recovery operations, the microchannels of the biochip are docked with the microchannels of the carrier to allow the solution to pass through. The embryos are restricted within the microchannels of the carrier by the grid structure and will not float out, enabling the orderly delivery or removal of cryoprotectants and effectively improving the efficiency of embryo / egg cryopreservation or thawing and recovery operations. Brief Description of the Drawings

[0013] The drawings described herein are used to provide a further understanding of the present invention and form a part of this application. The schematic embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute an improper limitation to the present invention. In the drawings:

[0014] Figure 1 is a three-dimensional schematic diagram of the device according to an embodiment of the present invention;

[0015] Figure 2 is a top view of the device according to an embodiment of the present invention;

[0016] Figure 3 is Figure 2 a schematic cross-sectional view taken along line A-A in

[0017] Figure 4 is a top view of the chip according to an embodiment of the present invention;

[0018] Figure 5 is Figure 4 a schematic cross-sectional view taken along line B-B in

[0019] Figure 6 is Figure 5 a partial enlarged view of C in

[0020] Figure 7 is Figure 6 a partial enlarged view of D in

[0021] Figure 8 is a schematic diagram of a glass capillary passing through the grid structure according to an embodiment of the present invention. Detailed Description of the Embodiments

[0022] To make the objectives, technical solutions, and advantages of the present invention clearer, the technical solutions of the present invention will be clearly and completely described below in conjunction with specific embodiments of the present invention and the corresponding drawings. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of them. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the scope of protection of the present invention.

[0023] The following will detail the technical solutions provided by each embodiment of the present invention with reference to the drawings.

[0024] According to an embodiment of the present invention, a device for cryopreservation or thawing and recovery of biological tissues is provided. Refer to Figure 1 , the device includes a biochip 1 and a carrier 2. Among them, the biochip 1 is generally in a cuboid structure, and a microchannel 11 (first microchannel) is provided inside the biochip 1. Solution inlets 12 and a solution outlet 13 are respectively provided at both ends of the microchannel 11 and are communicated with it. The solution inlet 12 is arranged on the upper surface of the biochip 1, and the solution outlet 13 is arranged on the lower surface of the biochip 1. The solution can enter the microchannel 11 through the solution inlet 12 and leave the microchannel 11 through the solution outlet 13. According to an embodiment of the present application, the biochip 1 is provided with two solution inlets 12. One of the solution inlets can be filled with a balanced solution, a vitrification cryoprotectant (for cryopreservation application), a thawing solution, a dilution solution, a cleaning solution (for thawing application), and the other solution inlet can be filled with a basic culture solution (BS). By controlling the pressure ratio of the two solution inlets, the concentration of the solution flowing into the microchannel 11 can be controlled. For example, if the pressure ratio of the balanced solution to the basic culture solution is 50:50, then the concentration of the solution flowing into the microchannel 11 and contacting the embryo is 50% of the balanced solution. In some embodiments of the present application, the microchannel 11 is arranged in a serpentine shape inside the biochip 1. In the limited internal space of the biochip 1, the serpentine arrangement can maximize the length of the microchannel 11 as much as possible, so that the solutions filled through the two solution inlets can be fully mixed in the microchannel 11.

[0025] The carrier 2 is generally in a long strip structure, and it includes a front-end thin sheet and a rear-end carrier rod. The front-end thin sheet of the carrier 2 can be made of a plastic material with uniform thickness, transparent material, biocompatibility, and good heat transfer performance, ensuring the applicability to holding embryos and the heat transfer speed during subsequent cryopreservation. The rear-end carrier rod of the carrier 2 can also be called a handle or a hand-held part, which is the position directly contacted and operated by the operator's hand. Only the front-end thin sheet part of the carrier is shown in the drawings in the present application, and the rear-end carrier rod part of the carrier 2 is not shown as a key part.

[0026] The interior of the carrier 2 has microchannels 21 (second microchannels), where the width of the microchannels 21 is 0.5 mm, the height is 0.2 - 0.5 mm, and the length is 20 mm. At both ends of the microchannels 21, a solution inlet 22 and a solution outlet 23 communicating therewith are respectively provided. The solution inlet 22 has a flat or smooth opening for facilitating fitting with the biochip 1, and the width of the opening can be 1 mm. At the upper and lower openings of the solution outlet 23, grid-like structures 25 are respectively provided. Or rather, the solution outlet 23 has a through-hole, and the diameter of the through-hole can be 0.5 - 1 mm. The grid-like structure 25 includes a plurality of grid bars, and the slit spacing between every two of these grid bars is less than the diameter of the biological tissue and greater than the diameter of the solution molecules, where the slit spacing can be 0.05 - 0.08 mm. In this way, the grid-like structure 25 can confine the biological tissue in the microchannels 21, preventing the biological tissue from floating out of the solution outlet 23. Among them, the biological tissue can be biological materials such as embryos, eggs, cells, etc., and the present application does not limit this.

[0027] When performing cryopreservation or thawing and recovery operations, the carrier 2 is closely attached below the biochip 1. The solution outlet 13 of the biochip 1 can be docked with the solution inlet 22 of the carrier 2, that is, the solution outlet 13 covers the solution inlet 22, and the solution flowing out through the solution outlet 13 can enter the microchannels 21 through the solution inlet 22.

[0028] In the embodiment of the present application, the grid bars of the grid-like structure 25 are elastic, and the grid-like structure 25 can deform when contacting an external object (such as a glass capillary) so that the external object passes through the grid-like structure from the slit between two grid bars. When using the glass capillary 3 to put an embryo into the carrier 2, with reference to Figure 8 After the glass capillary 5 touches the grid-like structure 25, the grid-like structure 25 will deform. Continuing to insert the glass capillary downward, the glass capillary 5 can then pass through the grid-like structure 25 and enter the microchannels 21, and the embryo in the glass capillary 5 is put into the microchannels 21. After the embryo is put in, when the glass capillary 5 leaves the microchannels 21, the grid-like structure 25 returns to its original state. Since the slit spacing of the grid-like structure 25 is less than the diameter of the embryo, the embryo can be limited in the microchannels 21 and will not leave the carrier 2 along with the flow of the liquid.

[0029] When performing the operation process of cryo-application, first place the biochip above the carrier and press it tightly. It should be noted that the solution outlet 13 on the lower surface of the chip needs to be docked with the solution inlet 22 of the carrier microchannel. Then, use a glass capillary to pass through the grid structure of the carrier to place the embryo in the groove of the carrier. Next, use a biochip solution propulsion instrument (such as a syringe pump or a pneumatic pump) to slowly pour the equilibration solution into the biochip from the solution inlet. By controlling the pressure ratio of the equilibration solution and the basic culture medium, the concentration of the cryoprotectant in the equilibration solution flowing to the embryo can be slowly and orderly increased, reducing the osmotic pressure damage to the embryo. After 10 minutes, the vitrification solution is poured into the chip. When reaching the microchannel outlet of the carrier, the cryoprotectant in the solution can diffuse into the embryo, achieving the purpose of cryoprotectant delivery and replacing the existing manual operation scheme. Since the outlet is provided with a grid structure, when the solution flowing into the carrier microchannel finally flows out from the outlet, the embryo will not leave the carrier along with the liquid flow. After the treatment is completed, the chip and the carrier can be directly put into liquid nitrogen for freezing; or in order to save space in the liquid nitrogen tank, the carrier and the chip can be separated first, and then the carrier can be put into liquid nitrogen for freezing.

[0030] When performing the thawing application operation, first prepare a plate of thawing solution preheated to 37 degrees, then take the carrier out of the liquid nitrogen, and immerse the transparent thin sheet part at the front end of the carrier into the thawing solution preheated to 37 degrees. After 3 - 5 seconds, the user places the chip above the carrier and presses it tightly. It should be noted that the outlet on the lower surface of the chip needs to be docked with the inlet of the carrier microchannel. Then, use a biochip solution propulsion instrument (such as a syringe pump or a pneumatic pump) to slowly pour the thawing solution into the chip from the solution inlet. By controlling the pressure ratio of the thawing solution and the washing solution, the concentration of the cryoprotectant in the thawing solution flowing to the embryo can be slowly and orderly decreased, reducing the osmotic pressure damage to the embryo. Since the dilution solution can be obtained by mixing the thawing solution and the washing solution, the entire thawing process can be completed by only adjusting the pressure ratio of the thawing solution and the washing solution. After about 10 minutes, the pressure ratio of the thawing solution and the washing solution drops to 0, and the cryoprotectant in and near the embryo is taken away and removed by the thawing solution, achieving the purpose of thawing and recovery and replacing the existing manual operation scheme. Since the outlet is provided with a grid structure, when the solution flowing into the carrier microchannel finally flows out from the outlet, the embryo will not leave the carrier along with the liquid flow. Finally, the user uses a glass capillary to pass through the grid structure of the carrier to recover the embryo from the microchannel and place it in a culture dish for continued cultivation.

[0031] Through the above embodiments of the present application, when performing cryopreservation or thawing and recovery operations, the grid structure restricts the embryo in the microchannel of the carrier and cannot float out, so that the delivery or removal of the cryoprotectant can be carried out orderly, effectively improving the efficiency of embryo cryopreservation or thawing and recovery operations.

[0032] The above are only embodiments of the present invention and are not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and changes. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included within the scope of the claims of the present invention.

Claims

1. A device for cryopreservation or thawing and recovery of biological tissues, characterized in that, Comprising: A biochip, the biochip having a first microchannel for the flow of a solution, the first microchannel including a first solution inlet and a first solution outlet, the first solution inlet being provided on the upper surface of the biochip, and the first solution outlet being provided on the lower surface of the biochip; A carrier, the carrier including a front-end thin sheet and a rear-end carrier rod, the front-end thin sheet of the carrier including a second microchannel provided with a second solution inlet and a second solution outlet, grid-like structures being respectively provided at the upper and lower openings of the second solution outlet, and the grid-like structures restricting the biological tissue in the second microchannel; Wherein, when performing cryopreservation or thawing and resuscitation operations, the carrier is closely attached below the biochip, and the second solution inlet of the carrier is docked with the first solution outlet of the biochip.

2. The device according to claim 1, characterized in that The number of the first solution inlets is two.

3. The device according to claim 1, characterized in that The first microchannel is serpentine.

4. The device according to claim 1, wherein The grid-like structure includes a plurality of grid bars, and the distance between every two grid bars is less than the diameter of the biological tissue and greater than the diameter of solution molecules.

5. The device according to claim 1, characterized in that, The first solution outlet of the biochip covers the second solution inlet of the carrier.

6. The device according to claim 5, characterized in that The biochip does not cover the second solution outlet of the carrier.

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