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Cell cryoprotectant and application thereof

A cryopreservation solution and cell technology, applied in the field of cell cryopreservation solution, can solve the problems of easily damaged cell membrane permeability, loss of cell content, spatial structure disorder, etc., to achieve stability, avoid cell damage, and maintain intact functions Effect

Inactive Publication Date: 2019-05-14
GUANGZHOU YIYANG BIO TECH CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, dehydration of the cells increases the local electrolyte concentration, changes the pH value, and denatures some proteins due to the above reasons, causing the internal space structure of the cell to be disordered. Causes damage, mitochondrial swelling, loss of function, and energy disturbance
The lipoprotein complex on the cell membrane is also easily destroyed, causing changes in the permeability of the cell membrane, resulting in the loss of cell contents

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] [Example 1] Application of cryopreservation solution in fibroblasts

[0028] An embodiment of the cell cryopreservation solution of the present invention, the cryopreservation solution is composed of the following components: 9% dimethyl sulfoxide, 0.3% trehalose, 1% human albumin, 10% horse serum , and the balance was DMEM medium.

[0029] The specific operation is: add 1% human albumin and 10% horse serum to DMEM medium to obtain solution I; add 9% dimethyl sulfoxide and 0.3% trehalose to DMEM medium to obtain solution II. Digest and centrifuge the fibroblasts that have been cultured to a period suitable for cryopreservation to collect the cells to obtain cell pellets; slowly add solution I to the cells and mix the cells; slowly drop solution II into the cell suspension and mix well cell suspension. According to the concentration of 1×10 6 ~5×10 6 Cell suspensions per tube, divided into cell suspensions, placed in a -20°C refrigerator for 2 hours, then placed at -...

Embodiment 2

[0031] [Example 2] Application of cryopreservation solution in umbilical cord mesenchymal stem cells

[0032] An embodiment of the cell cryopreservation solution of the present invention, the cryopreservation solution is composed of the following components: 10% dimethyl sulfoxide, 0.5% trehalose, 5% human albumin, 15% horse serum , and the balance was DMEM medium.

[0033] The specific operation is: add 5% human albumin and 15% horse serum to DMEM medium to obtain solution I; add 10% dimethyl sulfoxide and 0.5% trehalose to DMEM medium to obtain solution II. The umbilical cord mesenchymal stem cells that have been cultured to a period suitable for cryopreservation are digested with trypsin and centrifuged to collect the cells to obtain cell pellets; slowly add the obtained solution I to the cells and mix the cells; slowly drop solution II into the cell suspension , and mix the cell suspension. According to the concentration of 1×10 6~5×10 6 Pack the cell suspension in one...

Embodiment 3

[0035] [Example 3] Application of cryopreservation solution in immune cells

[0036] An embodiment of the cell cryopreservation liquid of the present invention, said cryopreservation liquid is made up of the composition of following concentration: 10% dimethyl sulfoxide, 0.4% dextran, 5% human serum albumin, and balance is Serum-free medium.

[0037] The specific operation is: adding 5% human serum albumin to serum-free medium to obtain solution I; adding 10% dimethyl sulfoxide and 0.4% trehalose to DMEM medium to obtain solution II. Centrifuge the immune cells cultured to the logarithmic growth phase, such as NK cells, CIK cells, DC cells, CART cells, etc., to collect the cells to obtain cell pellets; slowly add solution I to the cells and mix the cells; slowly add solution II Drop into the cell suspension and mix the cell suspension. According to the concentration of 1×10 6 ~5×10 6 Cell suspensions per tube, cell suspension; store in -20°C refrigerator for 2 hours, then ...

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PUM

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Abstract

The invention relates to the field of cells, particularly to a cell cryoprotectant and application thereof. The cell cryoprotectant is composed of mixtures of permeable protectant, non-permeable protectant, energy supply agent and the like and can be applied to cryopreservation of adherent cells, suspension cell and clinic application cells. The survival rate of cells recovered and cultured in thecell cryoprotectant can reach up to 98%, and detected various indexes of the cells show that the cells are functionally heathy, so that the cell cryoprotectant can well maintain the physiological functions of the cells and avoid mutation, tumorigenicity and the like. Meanwhile, the cell cryoprotectant is simple in preparation processes, easy to operate, wide in application range and capable of facilitating research and clinical popularization and application.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to a cell cryopreservation solution and its application. Background technique [0002] In the process of subculture and daily maintenance of cell culture, once the cells leave the living body and start primary culture, its various biological characteristics will gradually change and will continue to change with the increase in the number of passages and changes in in vitro environmental conditions . Storing cells in liquid nitrogen can temporarily remove the cells from the growth state and preserve their cellular characteristics, so that the cells can be revived for experiments when needed. Moreover, moderately preserving a certain amount of cells can prevent the cells from being lost due to contamination of the cells being cultured or other accidents, which plays a role in cell preservation. [0003] Cell cryopreservation methods often have problems such as low cell recovery...

Claims

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Application Information

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IPC IPC(8): A01N1/02C12N5/071
Inventor 夏黎明夏建川万阳杨思诗王亮
Owner GUANGZHOU YIYANG BIO TECH CO LTD
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