Freezing and thawing of MDCK cell

A cell and cryopreservation technology, applied in animal cells, vertebrate cells, urinary tract/kidney cells, etc., can solve the problems of surface antigen loss, low MDCK cell viability, affecting experimental results, etc., to achieve good cell viability and maintenance. Cell viability, the effect of ensuring cell quality

Inactive Publication Date: 2016-10-12
哈尔滨市疾病预防控制中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This is a relatively common method of cell cryopreservation at present. However, the operation of using the above-mentioned cryopreservation solution for cryopreservation is carried out by a relatively conventional method, which makes the survival rate of cryopreserved MDCK cells low after recovery, and the surface antigen is seriously lost. , affecting the experimental results

Method used

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  • Freezing and thawing of MDCK cell

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Experimental program
Comparison scheme
Effect test

specific Embodiment approach 1

[0018] Specific embodiment one: the cryopreservation of the MDCK cell of the present embodiment, it is carried out according to the following steps:

[0019] 1. Take the MDCK cells that have been subcultured to the logarithmic phase, digest and culture the cells, centrifuge, remove the supernatant, put them in an ice bath, add the cryopreservation solution to resuspend the cells, until the cell density reaches 5×10 6 / mL, to obtain the cell freezing solution; among them, the freezing solution was added dropwise along the wall of the cryopreservation tube with a dropper, and the dropping rate was: add dropwise at a rate of 0.2 mL / min for the first 3 minutes, and drop at a rate of 0.5 mL / min for 3 to 5 minutes. Add dropwise, and add dropwise at a rate of 0.75mL / min for the rest of the time;

[0020] 2. First lower the freezing tube to 4°C, then drop it to -8°C at a rate of 1.5°C / min, then take it out and put it in -10°C, then drop it to -20°C at a rate of 1.0°C / min, and then let...

specific Embodiment approach 2

[0022] Specific implementation mode two: the difference between this implementation mode and specific implementation mode one is: 1×10 7 10 mL of freezing solution was added dropwise to the cell density per mL. Others are the same as in the first embodiment.

specific Embodiment approach 3

[0023] Embodiment 3: This embodiment is different from Embodiment 1 in that: before the MDCK cells are frozen, the cell culture medium is replaced, and then the cells are digested and cultured. Others are the same as in the first embodiment.

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Abstract

The invention relates to freezing and thawing of MDCK cell, and relates to freezing and thawing of the MDCK cell. The freezing method comprises the following steps: by restricting the addition rate of the frozen stock solution in mixing of a frozen stock solution and the cell, cell damage is prevented, by adjusting the cooling rate, generation of ice crystal in the cell can be prevented, and cell death is lead. In cell thawing, thawing can be rapidly carried out, and integrity and survival rate of the cell can be guaranteed. The method can better keep the MDCK cell vitality. The cell vitality after thawing can reach more than 95%, the cell quality is guaranteed, and long-term viability of the cell can be guaranteed.

Description

technical field [0001] The invention relates to freezing storage and recovery of cells. Background technique [0002] The cryopreservation of cells is a complex problem plaguing the field of cryobiology research. The position, structure and function of cell junctions in tissues are special, so their response to freezing injury and the protection mechanism of freezing injury should also have their particularity. Therefore, it is necessary to systematically observe and study the characteristics and rules of cell junction damage in cryopreservation, establish targeted and effective cryopreservation technical solutions, and improve and ensure the cryopreservation effect of tissues and organs. Studies have shown that cell junctions may be one of the targets of various damage factors during cryopreservation, leading to damage and destruction of tissue structures. First of all, through the transcellular membrane protein, the cell connects with adjacent cells or matrix, and connec...

Claims

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Application Information

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IPC IPC(8): A01N1/02C12N5/071
CPCA01N1/0221C12N5/0686
Inventor 张崇华任莉娜魏丽娜
Owner 哈尔滨市疾病预防控制中心
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