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Biological material vitrified frozen vector and preparation method thereof

A technology of vitrification and biomaterials, applied in the field of biomedicine, can solve the problems of difficult temperature control of the heating device, inability to elongate the straw, easy breakage of the straw, etc., to prevent the loss of frozen specimens, and the production method is simple and easy , Easy to take and put

Inactive Publication Date: 2009-03-18
PEKING UNIV THIRD HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there are various problems in the production and use of the existing vitrification carriers, which affect the efficiency of cryopreservation and the development of vitrification technology.
Open straws or closed straws have the following problems: the temperature of the heating device (alcohol lamp or hot plate) is not easy to control during the production process, the straw cannot be elongated if the temperature is slightly lower, and the straw is easy to be stretched if the temperature is higher. It is easy to be damaged by external force and damaged during storage, resulting in the loss of frozen specimens
Electron microscope copper mesh, Cryoloop, Cryotop are easy to lose specimens during freezing and storage
Therefore, the above-mentioned cryocarriers have poor feasibility in clinical application and promotion, and are not widely accepted.
Cryoleaf is currently the most widely used freezing carrier in clinical practice, but it is expensive (about 200 yuan each), and it is difficult to be widely used in scientific research

Method used

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  • Biological material vitrified frozen vector and preparation method thereof
  • Biological material vitrified frozen vector and preparation method thereof
  • Biological material vitrified frozen vector and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Embodiment 1, the preparation of biological material vitrification freezing carrier

[0045] 1. Preparation of materials

[0046] Commercially available injection needles (18 gauge); commercially available syringe needle covers (18 gauge);

[0047] Straw (0.25ml): French I.M.V company.

[0048] 2. Preparation of biomaterial vitrification cryocarriers

[0049] 1) Use pliers to cut the front half of the injection needle into a flat opening to obtain a flat injection needle;

[0050] 2) Boil the double distilled water with an electric stove;

[0051] 3) Clamp both ends of the straw with a vascular clamp;

[0052] 4) Hold the vascular forceps, immerse the straw in the boiling water bath, and pull the straw to both ends to 4 times the length of the original pipe to obtain an open-type drawn thin straw (outer diameter 1.0mm, inner diameter 0.8mm);

[0053] 5) Lift the open-type drawn thin straw from the liquid surface, and maintain the original pulling force for a few se...

Embodiment 2

[0061] Embodiment 2, the application of biological material vitrification freezing carrier

[0062] The cryocarrier prepared in Example 1 was sterilized by fumigation with ethylene oxide. Fifty-three mouse mature oocytes were vitrified with sterilized freezing carriers. After freezing for 1 day, the oocytes were thawed and their survival was detected. Survival criteria for resuscitated oocytes are as follows: oocytes have good refractive properties, are transparent, have no obvious shrinkage of cytoplasm, and have complete zona pellucida and cell membranes.

[0063] Equilibrium solution (EM): 10% ethylene glycol + 10% dimethyl sulfoxide + 10% fetal bovine serum + DPBS;

[0064] Vitrification solution (VM): 20% ethylene glycol + 20% dimethyl sulfoxide + 10% fetal bovine serum + DPBS;

[0065] Thawing solution (T1 solution): 0.1M sucrose + 10% fetal bovine serum + DPBS;

[0066] Thawing solution (T2 solution): 0.5M sucrose + 10% fetal bovine serum + DPBS;

[0067] Thawing so...

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Abstract

The invention discloses a biomaterial vitrified freezing vector and a preparation method thereof. The biomaterial vitrified freezing vector provided by the invention comprises a syringe needle (1) and a wheat flake (2), wherein the syringe needle (1) consists of a needle (1-1) and a needle seat (1-2); the wheat flake (2) is an open finely-drawn straw; one end (2-1) of the wheat flake (2) is sleeved in an inner cavity of the needle (1-1); and the straw wall on the other free end (2-2) of the wheat flake (2) is partially lost, and the cross section of the residual straw wall is one fifth to one third of the circumference. The invention also provides the preparation method for the biomaterial vitrified freezing vector. The biomaterial vitrified freezing vector has the advantages of simple and convenient operation, short time consumption, low cost and so on, is suitable to be used by genocentric labs of various hospitals and various research units, and has important significance in promoting the development of the vitrified freezing technology in the medical field and the cryopreservation research on ovum, embryo, ovarium tissues and so on.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to a biological material vitrification freezing carrier and a preparation method thereof. Background technique [0002] At present, cryogenic freezing technology is widely used in the field of assisted reproduction. Vitrification technology uses a tiny cryogenic tool to transform the high-concentration cryoprotectant solution from a liquid to a glass-like solid with a high viscosity during an extremely rapid cooling (>2000°C / min) process, preventing cells from The formation of internal ice crystals can achieve the effect of cryoprotection, thereby significantly improving the viability and development ability of cells / tissues after freezing. Vitrification technology has the advantages of simplicity, high efficiency, time saving, safety, and low cost. In recent years, the application of vitrification technology in the cryopreservation of human eggs and embryos has gradually increased. Th...

Claims

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Application Information

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IPC IPC(8): C12M1/24
CPCC12M45/22
Inventor 乔杰闫丽盈严杰刘平廉颖
Owner PEKING UNIV THIRD HOSPITAL
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