Serum-free cell cryopreservation liquid

A cryopreservation, serum-free technology, applied in the biological field, can solve the problems of great difference in quality, cell influence, animal damage, etc., and achieve the effect of ensuring reliability, low pollution risk, and reducing pollution.

Inactive Publication Date: 2016-07-27
TIANJIN ZHONGAO TIANYUAN TECH DEV CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the prior art, the method of mixing serum and DMSO is used to freeze the cells. Each time the cells are frozen, the serum needs to be thawed first, and the survival rate of the cells after freezing and resuscitation is relatively low, especially some relatively fragile cells, and the resuscitation It is inconvenient to use after changing the medium; due to different animals, different serum origins and batch numbers, the quality of each batch varies greatly, and its components cannot be kept consistent; mycoplasma and viruses may be brought into the samples, which may have a potential impact on the cells and may cause The experiment failed or the experimental results were unreliable; the cost of serum is high, and the price of each 500ml serum can be as high as 3000-4000 yuan; the collection of serum in vivo has certain damage to animals

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] First, take the A549 cells in the logarithmic growth phase, digest the cells growing in a monolayer with trypsin, and transfer the cells growing in suspension directly to a 15ml centrifuge tube; then, centrifuge at 1000rpm for 5min; then, remove trypsin and old cells Add an appropriate amount of the above-mentioned serum-free cell freezing medium to the culture medium, blow gently with a pipette to arrange the cells evenly, count them, and adjust the final density of the cells in the freezing medium to 5×10 6 cells / ml; secondly, divide the cells into cryopreservation tubes, 1.5ml per tube; finally, place the cryopreservation tubes in liquid nitrogen at minus 100°C and store them for 30 days. After 30 days, the cryopreservation tube in liquid nitrogen was taken out, put into warm water at 35°C for cell recovery, and then the state of A549 cells, number of surviving cells, and cell adhesion were observed under a microscope.

Embodiment 2 to 3

[0020] The A549 cells in Example 1 were replaced with PC3 cells and MB231 cells respectively, and the rest remained unchanged.

Embodiment 4 to 6

[0022] The serum-free cell freezing solution in Examples 1 to 3 was replaced with a conventional freezing solution, and the rest remained unchanged.

[0023] The observed results are shown in Table 1.

[0024] Table 1

[0025] Example

[0026] It can be clearly seen from the above table that the cells using this serum-free cryopreservation solution have a high survival rate after thawing, and the cells adhere to the wall in a good state, and there is no need to replace the cryopreservation solution; while the survival rate of cells using a conventional cryopreservation solution is extremely low after recovery. Many, many cells are not adhered to the wall, and the cryopreservation solution needs to be replaced.

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PUM

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Abstract

The invention discloses serum-free cell cryopreservation liquid and belongs to the biotechnology field.The serum-free cell cryopreservation liquid is used for solving the problems that existing cryopreservation liquid is inconvenient to use, high in cost and low in cell survival rate.The serum-free cell cryopreservation liquid contains, by volume, 80%-89% of serum-free medium and 11%-20% of a serum and DMSO substitute.The serum and DMSO substitute contains, by volume, 30% of glycerol, 20% of chitosan and 50% of alginic acid.According to the technical scheme, use is convenient, cost is low, and cells are high in survival rate after recovery.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a serum-free cell cryopreservation solution. Background technique [0002] Cell cryopreservation and recovery are one of the important technologies in cell culture technology, and cell cryopreservation is a method to preserve cultured cells. The basic principle of cell cryopreservation and recovery is slow freezing and quick thawing, which can preserve cell viability to the greatest extent. [0003] Cell cryopreservation solution is a solution that must be used for cell cryopreservation. Its function is to suspend cells that need to be frozen in the cryopreservation solution, supply the nutrients necessary for cell life metabolism, and prevent or reduce the impact of frozen ice crystals on cells. damage effect. In the prior art, the method of mixing serum and DMSO is used to freeze the cells. Each time the cells are frozen, the serum needs to be thawed first, and the survival rate ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221
Inventor 张顺锋
Owner TIANJIN ZHONGAO TIANYUAN TECH DEV CO LTD
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