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 and a carrier; the biochip includes a solution inlet and a solution outlet; a permeable membrane is provided at the bottom of the biochip, and a microchannel is formed between the biochip and the permeable membrane; the carrier has a groove, and the groove contains biological tissues; wherein, the carrier is closely attached below the chip, the permeable membrane covers the groove and confines the biological tissues in the groove, and the solution in the microchannel diffuses through the permeable membrane into the groove. By means of the present invention, the delivery or removal of cryoprotectant can be carried out orderly, 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 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 freezing 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 freezing or thawing.

[0005] According to an embodiment of the present invention, a device for cryopreservation or thawing and recovery of biological tissues is proposed, which includes: a biochip and a carrier; the biochip includes a solution inlet and a solution outlet; a permeable film is provided at the bottom of the biochip, and a microchannel is formed between the biochip and the permeable film; the carrier has a groove, and a biological tissue is carried in the groove; wherein, the carrier is closely attached below the chip, the permeable film covers the groove and confines the biological tissue in the groove, and the solution in the microchannel diffuses through the permeable film into the groove.

[0006] Wherein, the permeable film is provided with a plurality of micropores, the diameter of the micropores is smaller than the diameter of the biological tissue, and the diameter of the micropores is larger than the diameter of the solution molecules.

[0007] Wherein, the permeable film is a perforated film, a grid-like film, a dialysis film, or a water-soluble film.

[0008] Wherein, the biochip includes two solution inlets and one solution outlet.

[0009] Wherein, the solution inlet and the solution outlet of the biochip are respectively communicated with the microchannel.

[0010] Wherein, the carrier includes a front-end thin sheet and a rear-end carrier rod, and the groove is provided on the front-end thin sheet.

[0011] According to the technical solution of the present invention, by arranging a permeability film that only allows the solution to pass through and does not allow biological tissue to pass through under the biochip, during the cryopreservation or thawing and recovery operations, the embryo / egg is restricted by the permeability film in the groove and cannot float out, and the cryoprotectant or thawing solution flows in the microchannel and passes through the permeability film into the groove to contact the embryo / egg, 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. Brief Description of the Drawings

[0012] 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:

[0013] Figure 1 is a top view schematic diagram of a biochip and a carrier according to an embodiment of the present invention;

[0014] Figure 2 is along Figure 1 a schematic cross-sectional view of the line A-A in

[0015] Figure 3 is Figure 2 an enlarged view of the partial B in Detailed Description of the Embodiments

[0016] 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 the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

[0017] The following will detail the technical solutions provided by the embodiments of the present invention with reference to the drawings.

[0018] According to an embodiment of the present invention, a device for cryopreserving or thawing and recovering biological tissue is provided. Referring to Figures 1 to 3 , the device includes: a biochip 1 and a carrier 2, wherein the biochip 1 is generally in a cuboid structure and may include two solution inlets 11 and 12 and a solution outlet 13. In some embodiments, the number of solution inlets of the biochip 1 can be set according to requirements. 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 illustrative, and the important thing is that the solutions flowing into the biochip 1 from different inlets can flow to the vicinity of the embryo in sequence.

[0019] The carrier 2 is generally in a long strip structure, which 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 its applicability to holding embryos and the heat transfer speed during subsequent freezing. 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. In the drawings of this application, only the front-end thin sheet part of the carrier is shown, and the rear-end carrier rod part of the carrier 2 is not shown as a key part. At the front end of the carrier 2 (front-end thin sheet), there is a groove 21, which is used to hold the biological tissue to be processed and the related solution for protecting the biological tissue. The surface of the groove 21 can be circular. Among them, the biological tissue can be biological materials such as embryos, eggs, cells, etc., and this application does not limit this. In the embodiment of this application, the thickness of the carrier 2 can be 0.25 - 1 mm, the depth of the groove 21 can be 0.2 - 0.8 mm, and the diameter of the groove 21 can be 0.2 - 1 mm.

[0020] A permeability thin film 3 is provided at the bottom (or lower surface) of the biochip 1. The permeability thin film 3 seals the bottom of the biochip and forms a microchannel 4, and the microchannel 4 is connected to a solution inlet and a solution outlet. The permeability thin film 3 covers the upper part of the carrier 2 and seals the groove 21. In the embodiment of this application, the permeability thin film 3 has a plurality of micropores, the diameter (pore size) of which is smaller than the diameter of the biological tissue but larger than the diameter of the solution (freezing solution or thawing solution) molecules. During the freezing and / or thawing operation process, the permeability thin film 3 can only allow the solution to pass through, and does not allow the biological tissue to pass through, so that the biological tissue is restricted within the groove 21 and will not float out of the groove 21. Among them, the permeability thin film 3 can be a perforated film, a grid-like thin film, a dialysis film, or a water-soluble thin film, etc., which are thin films with selective permeability.

[0021] When performing the freezing application operation process, place the biochip 1 on the top of the carrier 2 and press it tightly. Among them, the embryo is carried in the groove 21 of the carrier 2. A microchannel 4 that only allows the solution to pass through is formed between the biochip 1 and the permeability thin film 3 provided at its bottom. Use a biochip solution pushing instrument (such as an injection pump or a pneumatic pump) to slowly pour the freezing solution into the interior of the biochip 1 from the solution inlet. The solution flows in the microchannel 4. When the solution flows to the position above the groove 21, the cryoprotectant in the solution can diffuse through the permeability thin film 3 into the embryo interior, achieving the purpose of delivering the cryoprotectant and replacing the existing manual operation scheme. Then the solution leaves the biochip 1 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 outlet of the biochip 1 to collect the flowing waste liquid (generally a solution of dozens to hundreds of microliters). Finally, the user separates the carrier 2 and the biochip 1, and puts the carrier 2 into liquid nitrogen for freezing.

[0022] When performing the thawing application operation, take the carrier 2 out of the liquid nitrogen environment, press the transparent thin film part at the front end of the carrier 2 onto the hot stage that has been preheated to 37 degrees, and then use the biochip 1 that has also been preheated to 37 degrees to press the transparent thin film part at the front end of the carrier 2. Since the carrier 2 is very thin, it can also reach a sufficient heating rate on the hot stage. After the biochip 1 and the carrier 2 are attached, a biochip solution pushing instrument (such as an injection pump or a pneumatic pump) can be used to slowly pour solutions such as thawing fluid into the interior of the biochip 1 from the solution inlet according to an automatic program. The solution flows in the microchannel 4. When the solution flows to the position above the groove 21, the cryoprotectant in and near the embryo diffuses through the permeability film 3 into the thawing fluid in the microchannel 4 and is carried away by the thawing fluid, achieving the purpose of thawing and recovery, and replacing the existing manual operation solution. Then the solution leaves the biochip 1 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 outlet of the biochip 1 to collect the flowing waste liquid (generally a solution of dozens to hundreds of microliters). Finally, the user separates the carrier 2 and the biochip 1, and then can use a glass capillary to take out the embryo in the groove 21 and place it in a culture dish for continued culture.

[0023] Through the embodiments of the present application, when performing cryopreservation or thawing and recovery operations, the biochip provided with a permeability film is attached to the carrier. The permeability film confines the embryo in the carrier groove and prevents it from floating out. The freezing fluid or thawing fluid flows in the microchannel and passes through the permeability film to enter the groove and contact the embryo. In this way, the delivery or removal of the cryoprotectant can be carried out in an orderly manner, and the efficiency of embryo cryopreservation or thawing and recovery operations can be effectively improved.

[0024] The above are only the embodiments of the present invention and are not used to limit the present invention. For those skilled in the art, the present invention can have various changes and modifications. 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 and a carrier; The biochip includes a solution inlet and a solution outlet; a permeable membrane is provided at the bottom of the biochip, and a microchannel is formed between the biochip and the permeable membrane. The solution inlet and the solution outlet of the biochip are respectively communicated with the microchannel; The carrier includes a front-end thin sheet and a rear-end carrier rod. The front-end thin sheet has a groove, and a biological tissue is carried in the groove; Wherein, the carrier is closely attached below the chip, the permeable membrane covers the groove and confines the biological tissue in the groove, and the solution in the microchannel diffuses through the permeable membrane into the groove.

2. The device according to claim 1, wherein The permeable membrane is provided with a plurality of micropores, the diameter of the micropores is smaller than the diameter of the biological tissue, and the diameter of the micropores is larger than the diameter of the solution molecules.

3. The device according to claim 1 or 2, characterized in that, The permeable membrane is one of a perforated membrane, a mesh-like membrane, a dialysis membrane, or a water-soluble membrane.

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

Images