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. The device according to an embodiment of the present invention includes: a biochip, which includes a solution inlet and a solution outlet; a carrier, which has a first groove, and the biological tissue is carried in the first groove; wherein, the carrier is closely attached below the biochip, and a microchannel passing through the first groove is formed between the biochip and the carrier, and only the solution can pass through the microchannel. Through the present invention, the delivery or removal of cryoprotectant can be carried out in an orderly manner, effectively improving the efficiency of embryo / egg cryopreservation or thawing and recovery operations.

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 the cryopreserved state to the normal metabolic state. The thawing and recovery process involves the process of 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] According to an embodiment of the present invention, a device for cryopreservation or thawing and recovery of biological tissues is provided, which includes: a biochip including a solution inlet and a solution outlet; a carrier having a first groove in which a biological tissue is carried; wherein, the carrier is closely attached to the lower side of the biochip, and a microchannel passing through the first groove is formed between the biochip and the carrier, and the microchannel only allows the solution to pass through.

[0006] Wherein, at least a part of the bottom of the biochip is an open area, the carrier is closely attached to the lower side of the biochip and closes the open area, and the height or width of the microchannel between the biochip and the carrier is smaller than the size of the biological tissue.

[0007] Wherein, the bottom of the biochip has a protrusion, and a microchannel including a solution micro-inlet and a solution micro-outlet is formed between the protrusion and the first groove, and the widths of the solution micro-inlet and the solution micro-outlet are smaller than the size of the biological tissue.

[0008] Wherein, the protrusion faces the first groove, and the length of the first groove is greater than the length of the protrusion.

[0009] Wherein, the carrier is further provided with a second groove whose length is greater than that of the first groove, and the first groove is arranged within the second groove; the bottom of the biochip has a protrusion, and the microchannel is formed between the protrusion and the second groove, and the height or width of the microchannel is smaller than the size of the biological tissue.

[0010] Wherein, the length of the protruding part is less than the length of the second groove and greater than the length of the first groove.

[0011] Wherein, the biochip includes two solution inlets and one solution outlet.

[0012] According to the technical solution of the present invention, the biochip is closely attached to the carrier, and a microchannel for only the solution to pass through is formed between the biochip and the carrier. During cryopreservation or thawing and recovery operations, the cryogenic fluid or thawing fluid flows through the microchannel and contacts the embryo / egg. The embryo / egg is restricted in the groove, so that the delivery or removal of the cryoprotectant can be carried out in an orderly manner, and the efficiency of the cryopreservation or thawing and recovery operation of the embryo / egg is effectively improved. Description of the Drawings

[0013] The drawings described herein are used to provide a further understanding of the present invention and constitute 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 1A is a three-dimensional schematic diagram of the device according to an embodiment of the present invention;

[0015] Figure 1B is Figure 1A the top view of the device in

[0016] Figure 1C is the sectional view along Figure 1A the line 1C-1C in

[0017] Figure 1D is Figure 1C the enlarged view of the local part in

[0018] Figure 2A is the top view of the device according to another embodiment of the present invention;

[0019] Figure 2B is the sectional view along Figure 2A the line 2B-2B in

[0020] Figure 2C is Figure 2B the enlarged view of the local part in

[0021] Figure 3A is the top view of the device according to still another embodiment of the present invention;

[0022] Figure 3B is the sectional view along Figure 3A the line 3B-3B in

[0023] Figure 3C is Figure 3B the enlarged view of the local part in. Detailed implementation manners

[0024] 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 embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without making creative efforts fall within the scope of protection of the present invention.

[0025] The following will detail the technical solutions provided by each embodiment of the present invention with reference to the drawings.

[0026] According to an embodiment of the present invention, a device for cryopreservation or thawing and recovery of biological tissues is provided. Referring to Figures 1A to 1D , the device includes: a biochip 11 and a carrier 12. The biochip 11 is generally in a cuboid structure. Part or all of the bottom (or lower surface) of the biochip is an open area, that is, the entire bottom of the biochip is open, or the bottom area of the biochip corresponding to the carrier is open. The biochip 11 may include two solution inlets 111 and 112 and a solution outlet 113. The number of solution inlets of the biochip can be adjusted according to actual needs. If multiple different cryoprotectant solutions need to flow through the embryo in sequence, multiple solution inlets can be set. The inlet channels shown in the figure are only for illustrative purposes. Importantly, the solutions flowing into the biochip from different inlets can flow to the vicinity of the embryo in sequence.

[0027] The carrier 12 is generally in a long strip structure and includes a front-end thin sheet and a rear-end carrier rod. The front-end thin sheet of the carrier 12 can be made of a plastic material with uniform thickness, transparent material, biocompatibility and good heat transfer performance to ensure the applicability to holding the embryo and the heat transfer speed during subsequent freezing. The rear-end carrier rod of the carrier 12 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 of the present application, and the rear-end carrier rod part of the carrier is not shown as a key part. At the front end of the carrier 12 (front-end thin sheet), there is a first groove 121 for holding the biological tissue to be processed and the related solution for protecting the biological tissue. The surface of the first groove 121 can be circular. Among them, the biological tissue can be biological materials such as embryos, eggs, cells, etc., and the present application does not limit this. In the embodiment of the present application, the thickness of the carrier can be 0.25 - 0.7 mm, the depth of the first groove can be 0.2 - 0.5 mm, and the diameter of the first groove can be 0.2 - 1 mm.

[0028] During the freezing or thawing operation, since all or part of the bottom area of the biochip 11 is open, when the biochip 11 is attached to the carrier 12, the carrier 12 closes the open area at the bottom of the biochip 11, forming a microchannel 13 that passes through the first groove 121 and only allows the solution to pass through. The height or width of the microchannel 13 is smaller than the diameter of the embryo / egg. Specifically, the microchannel 13 includes a solution micro-inlet 131, a solution micro-outlet 132, a solution inlet channel, and a solution outlet channel. The solution inlet channel is between the solution micro-inlet 131 and the first groove 121, and the solution outlet channel is between the first groove 121 and the solution micro-outlet 132. Moreover, the height or width of the solution micro-inlet 131, the solution micro-outlet 132, the solution inlet channel, and the solution outlet channel are equal (or approximately equal), and this height or width is smaller than the diameter of the embryo / egg. In this way, during the freezing and / or thawing operation, only the solution (freezing solution or thawing solution) can pass through the microchannel 13 and the embryo / egg cannot pass through, so that the embryo / egg is restricted within the first groove 121 and will not float out of the first groove 121.

[0029] When performing the cryopreservation operation, place the biochip 11 above the carrier 12 and press it tightly. The embryo is carried in the first groove 121 of the carrier. There is a microchannel that only allows the solution to pass through between the biochip 11 and the carrier 12. Use a biochip solution propulsion instrument (such as an injection pump or a pneumatic pump) to slowly pour the freezing solution into the interior of the biochip 11 from the solution inlet. When the solution flows through the microchannel 13 to the first groove 121, the cryoprotectant in the solution can diffuse into the embryo, achieving the purpose of delivering the cryoprotectant and replacing the existing manual operation scheme. Then the solution leaves the biochip through the solution outlet. The solution outlet can be connected to a hose to lead the waste liquid to a waste liquid collection tube, or a groove can be designed at the biochip outlet to collect the flowing waste liquid (generally a solution of dozens to hundreds of microliters). Finally, the user separates the carrier and the biochip and puts the carrier into liquid nitrogen for freezing.

[0030] When performing the thawing and resuscitation operation, the carrier 12 is taken out of the liquid nitrogen environment, and the transparent thin sheet part at the front end of the carrier 12 is pressed onto a heating table preheated to 37 degrees Celsius. Then, the biochip 11, which is also preheated to 37 degrees Celsius, is used to press the transparent thin sheet part at the front end of the carrier 12. Since the carrier 12 is very thin, it can reach a sufficient heating rate on the heating table. When the biochip 11 and the carrier 12 are in contact, a biochip solution pushing instrument (such as a syringe pump or a pneumatic pump) can be used to slowly pour solutions such as thawing solution into the biochip interior according to an automatic program through the solution inlet. When the solution flows through the microchannels to the grooves, the cryoprotectant in and around the embryo is removed by the thawing solution, achieving the purpose of thawing and resuscitation, replacing the existing manual operation scheme. The solution leaves the biochip through the solution outlet. The outlet can be connected to a hose to lead the waste liquid to a waste liquid collection tube, or a groove can be designed at the biochip outlet to collect the outflowing waste liquid (generally a solution of dozens to hundreds of microliters). Finally, the user separates the carrier and the biochip, and then can use a glass capillary to take out the embryo in the groove and place it in a culture dish for continued culture.

[0031] According to an embodiment of the present invention, there is also provided a device for cryopreservation or thawing and resuscitation of biological tissues. Refer to Figures 2A to 2C , the device includes: a biochip 21 and a carrier 22. The biochip 21 may include two solution inlets 211 and 212 and a solution outlet 213. The similarities between the biochip 21 and the carrier 22 and the biochip 11 and the carrier 12 in the above-described embodiments will not be elaborated here. Only the differences will be described below.

[0032] The bottom of the biochip 21 has a protrusion 215, and the protrusion 215 makes the bottom of the biochip 21 form a segmented structure. Among them, the protrusion 215 is opposite to the first groove 221. Generally, the length of the protrusion 215 is less than the length of the first groove 221. In the embodiment of the present application, a microchannel 23 that passes through the first groove 221 and can only allow a solution (freezing solution or thawing solution) to pass is formed between the protrusion 215 and the first groove 221. The width of the microchannel 23 is less than the diameter of the embryo / egg. Specifically, the microchannel 23 includes a solution micro-inlet 231 and a solution micro-outlet 232, where the widths of the solution micro-inlet 231 and the solution micro-outlet 232 are equal (or approximately equal), and the widths of the solution micro-inlet 231 and the solution micro-outlet 232 are less than the diameter of the embryo / egg. In this way, during the freezing and / or thawing operation, the microchannel 23 can only allow a solution (freezing solution or thawing solution) to pass and does not allow the embryo / egg to pass, so that the embryo / egg is restricted within the first groove 221 and will not float out of the first groove 221. For the specific freezing and thawing processes, please refer to the previous description and will not be elaborated here.

[0033] According to an embodiment of the present invention, there is also provided a device for cryopreservation or thawing and recovery of biological tissues. Refer to Figures 3A to 3C , the device includes: a biochip 31 and a carrier 32. The biochip 31 includes two solution inlets 311 and 312 and one solution outlet 313. The general structures of the biochip 31 and the carrier 32 are similar to those of the embodiments described above, which will not be elaborated here. Only the differences will be described below.

[0034] The carrier 32 includes a first groove 321 and a second groove 322. The first groove 321 is disposed within the second groove 322 and the first groove 321 can be located at the middle position of the second groove 322. In the embodiments of the present application, the length of the second groove 322 is greater than the length of the first groove 321, and the depth of the second groove 322 is less than the depth of the first groove 321. For example, the thickness of the carrier 32 is 0.25 - 0.7 mm, the depth of the first groove is 0.2 - 0.5 mm, and the diameter of the first groove is 0.2 - 1 mm. The depth of the second groove is 0.01 - 0.08 mm, and the length of the second groove is 2 - 10 mm.

[0035] The bottom of the biochip 31 has a protrusion 315, and the protrusion 315 makes the bottom of the biochip 31 form a segmented structure. Among them, the length of the protrusion 315 is less than the length of the second groove 322 and greater than the length of the first groove 321. For example, the length of the protrusion 315 is 1 - 6 mm. In the embodiments of the present application, a microchannel 33 that passes through the first groove 321 and can only allow a solution (cryoprotectant or thawing solution) to pass through is formed between the protrusion 315 and the second groove 322. The height or width of the microchannel 33 is less than the diameter of the embryo / egg. Specifically, the microchannel 33 includes a solution micro-inlet 331, a solution micro-outlet 332, a solution inlet channel, and a solution outlet channel. Among them, the solution inlet channel is between the solution micro-inlet 331 and the first groove 321, and the solution outlet channel is between the first groove 21 and the solution micro-outlet 332. And, the height or width of the solution micro-inlet 331, the solution micro-outlet 332, the solution inlet channel, and the solution outlet channel are equal (or approximately equal), and this height or width is less than the diameter of the embryo / egg. In this way, during the freezing and / or thawing operation, the microchannel 33 can only allow a solution (cryoprotectant or thawing solution) to pass through and does not allow the embryo / egg to pass through, so that the embryo / egg is restricted within the first groove 321 and will not float out of the first groove 321. For the specific freezing and thawing processes, please refer to the previous description, which will not be elaborated here.

[0036] Through the embodiments of the present application, during the cryopreservation or thawing and recovery operation, the embryo / egg can be restricted within the carrier groove, so that the delivery or removal of the cryoprotectant can be carried out in an orderly manner, and the efficiency of the cryopreservation or thawing and recovery operation of the embryo / egg is effectively improved.

[0037] 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, which includes a solution inlet and a solution outlet, and the bottom of the biochip has an open area communicating with the solution inlet and the solution outlet; A carrier, which includes a front sheet and a rear carrier rod, and the front end of the carrier has a first groove, and a biological tissue is carried in the first groove; Wherein, the carrier is closely attached below the biochip and closes the open area, and a microchannel passing through the first groove is formed between the biochip and the carrier, the microchannel is arranged between the solution inlet and the solution outlet of the biochip, and only the solution can pass through the microchannel.

2. The device according to claim 1, wherein The height or width of the microchannel between the biochip and the carrier is smaller than the size of the biological tissue.

3. The device according to claim 1, characterized in that The bottom of the biochip has a protrusion, and a microchannel including a solution micro-inlet and a solution micro-outlet is formed between the protrusion and the first groove, wherein the widths of the solution micro-inlet and the solution micro-outlet are smaller than the size of the biological tissue.

4. The device according to claim 3, characterized in that, The protrusion is opposite to the first groove, and the length of the first groove is greater than the length of the protrusion.

5. The device according to claim 1, characterized in that, The carrier is further provided with a second groove whose length is greater than that of the first groove, and the first groove is arranged within the second groove; the bottom of the biochip has a protrusion, and the microchannel is formed between the protrusion and the second groove, wherein the height or width of the microchannel is smaller than the size of the biological tissue.

6. The device according to claim 5, characterized in that, The length of the protrusion is smaller than the length of the second groove and greater than the length of the first groove.

7. The device according to claim 1, characterized in that, The biochip includes two solution inlets and one solution outlet.

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