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Adherent tumor cell cryopreservation method and improved cryopreservation solution formula

A technology of tumor cell and cryopreservation method, which is applied to the preservation, application, and animal husbandry of human or animal bodies. It can solve the problems of cumbersome operation process, low cell activity, and long operation time, and achieve simple preparation method and simple operation. , easy-to-operate effect

Inactive Publication Date: 2018-07-20
GUIZHOU UNIV
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  • Summary
  • Abstract
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Problems solved by technology

[0005] The purpose of the present invention is to: provide a method for cryopreservation of adherent tumor cells and an improved formula of cryopreservation solution to solve the problem that the traditional method of cryopreservation of adherent tumor cells takes a long time to experiment and requires centrifugation to remove the culture medium. It is cumbersome and causes great damage to cells, and the activity of cells after recovery is low.

Method used

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Effect test

Embodiment

[0017] This embodiment provides an improved cryopreservation solution formula, which includes: 30% fetal bovine serum, 20% dimethyl sulfoxide and 50% DMEM medium by weight.

[0018] The method for cryopreserving adherent tumor cells using the above formula is as follows:

[0019] First, configure the improved cell cryopreservation solution, take a sterile centrifuge tube (50ml), add 25ml DMEM medium into the centrifuge tube with a pipette, then slowly add 10ml DMSO while shaking, and finally add 15ml FBS, after the preparation is completed , aliquoted into 1ml / tube, and stored in a -20°C refrigerator.

[0020] When freezing the cells, first discard the old medium, add sterile PBS along the opposite side of the cells to wash twice, then add 1ml of trypsin, shake the cell culture flask gently in a cross shape for 5 seconds, and then suck up the trypsin with a Pasteur pipette , placed in a 37°C incubator and incubated for 3 minutes to digest the cells, then added 2ml of fresh me...

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Abstract

The invention provides an adherent tumor cell cryopreservation method and an improved cryopreservation solution formula. By configuring an improved cell cryopreservation solution, the intermediate centrifugation step is omitted, so that the experimental operation time is greatly shortened, the experimental consumables can be saved, the experimental efficiency is increased, and the problems that the traditional adherent tumor cell cryopreservation method is longer in experimental operation time, needs to remove the culture solution by means of centrifugation, and is cumbersome in operation process and larger in damage to the cells, the activity of the resuscitated cells is lower, and the like can be solved. The adherent tumor cell cryopreservation method and the improved cryopreservation solution formula belong to the technical field of biology.

Description

technical field [0001] The invention provides a method for cryopreserving adherent tumor cells and a formula of a cryopreservation solution, belonging to the field of biotechnology. technical background [0002] Factors such as selective subculture in the early stage of cell culture, senescence of limited cell lines, and genetic instability in continuous cell lines can promote the tendency of cell lines to mutate during continuous culture. In addition, accidents may also occur in the laboratory, such as a sudden power failure of the cell incubator, or even human contamination of cells during operation and cross-contamination between cell lines, etc. Therefore, cryopreservation of cells in experiments is of great significance. [0003] Ultra-low temperature cryopreservation is an effective method for long-term preservation of cells, which mainly plays a protective role by reducing the metabolic rate of cells. Liquid nitrogen (-196°C) preservation of cells is the most common ...

Claims

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Application Information

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IPC IPC(8): A01N1/02
CPCA01N1/0221A01N1/0226
Inventor 郭建军檀军田莹赵帅张书琪
Owner GUIZHOU UNIV
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