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Breast stem cell cryopreservation liquid, preparation method and cryopreservation method thereof

A technology of mammary gland stem cells and cryopreservation method, which is applied to the preservation, application, and animal husbandry of human or animal bodies, and can solve the problems of mammary gland stem cell damage, poor cryopreservation effect, and loss of function, so as to avoid toxic effects and maintain Multidirectional differentiation and self-renewal ability, and the effect of maintaining stemness

Inactive Publication Date: 2021-09-07
NINGXIA MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The most commonly used cryoprotectant in cell cryopreservation is dimethyl sulfoxide (DMSO), and too high a concentration of DMSO has a cytotoxic effect, and too low a concentration may lead to poor cryopreservation, resulting in damage and loss of function of mammary stem cells even death

Method used

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  • Breast stem cell cryopreservation liquid, preparation method and cryopreservation method thereof
  • Breast stem cell cryopreservation liquid, preparation method and cryopreservation method thereof
  • Breast stem cell cryopreservation liquid, preparation method and cryopreservation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0035] A breast stem cell lysery, and the freeze-containing mix is ​​mixed by a liquid and B liquid:

[0036] The volume ratio of the A liquid to the B liquid is 1: 1;

[0037] The A liquid is 100 parts by volume, including the following volume of components: 25 parts of glycerin and 75 calf serum;

[0038] The B liquid is 100 parts per scale, including the following components: 75 DMEM / F12 basic medium, 20 calf serum, 2 parts without vitamin A type B-27 (50x), 2.5 parts of heparin (2μg / ML), 0.4 parts of insulin (10 μg / mL), 0.04 recombinant basic fibroblast growth factor (10 ng / ml), 0.05 parts of epithelial cell growth factor (4 ng / ml) and 0.01 y-27632 (100 mM).

Embodiment 2

[0040] The preferred embodiment of the preferred embodiment of the present invention is provided with a mammary stem cell lysery, and the freeze-containing mix is ​​mixed by a liquid and a B liquid:

[0041] The volume ratio of a liquid and B liquid is 2: 1;

[0042] The A fluid is 100 parts per meter, including the following volume: 40 parts of glycerol and 60 calf serum;

[0043] The B liquid is 100 parts per scale, including the following components: 75 DMEM / F12 basic medium, 20 calf serum, 2 parts without vitamin A type B-27 (50x), 2.5 parts of heparin (2μg / ML), 0.4 parts of insulin (10 μg / mL), 0.04 recombinant basic fibroblast growth factor (10 ng / ml), 0.05 parts of epithelial cell growth factor (4 ng / ml) and 0.01 y-27632 (100 mM).

Embodiment 3

[0045] The preferred embodiment of the preferred embodiment of the present invention is provided with a mammary stem cell lysery, and the freeze-containing mix is ​​mixed by a liquid and a B liquid:

[0046] The volume ratio of a liquid and B liquid is 2: 1;

[0047] The A liquid is 100 parts by volume, including the following volume of components: 20 parts of glycerin and 80 calf serum;

[0048]The B liquid is 100 parts per scale, including the following components: 75 DMEM / F12 basic medium, 20 calf serum, 2 parts without vitamin A type B-27 (50x), 2.5 parts of heparin (2μg / ML), 0.4 parts of insulin (10 μg / mL), 0.04 recombinant basic fibroblast growth factor (10 ng / ml), 0.05 parts of epithelial cell growth factor (4 ng / ml) and 0.01 y-27632 (100 mM).

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Abstract

The invention discloses a breast stem cell cryopreservation liquid, which comprises a solution A and a solution B, wherein the volume ratio of the solution A to the solution B is (1-3):1, the solution A is prepared from the following components in parts by volume: 20 to 40 parts of glycerol and 60 to 80 parts of calf serum according to the total volume fraction of 100 parts, and the solution B is prepared from the following components in parts by volume: 75 to 80 parts of DMEM / F12 basal culture medium, 10 to 20 parts of calf serum, 2 parts of vitamin A-free B-27, 0.5 part of heparin sodium, 0.4 part of insulin, 0.04 part of recombinant basic fibroblast growth factors, 0.05 part of epidermal growth factors and 0.01 part of Y-27632. By using the cryopreservation liquid, the cryopreservation efficiency can be remarkably improved, the ice crystal damage during cell cryopreservation is reduced, so that the motility of resuscitated stem cells is improved, the original dryness of the stem cells can be maintained, and the multidirectional differentiation and self-renewal capabilities of the stem cells are maintained.

Description

Technical field [0001] The present invention belongs to the field of stem cells, which relates to a breast stem cell lysed solution, and the present invention also relates to the preparation method of the above freeze-stock solution, and also relates to the frozen deposit method of the above freezing liquid. Background technique [0002] Breast stem cells are adult stem cells with multi-directional differentiation and self-renewal capabilities, and play a crucial role in tissue self-steady-state maintenance and breast cancer. In laboratory research and clinical applications, the main storage method of breast stem cells is to store it in liquid nitrogen in liquid nitrogen in liquid nitrogen. The most commonly used refrigerant in cytometry is dimethyl sulfoxide (DMSO), while excessive concentrations of DMSO have cytotoxic effects, which may result in poorly frozen effect, resulting in breast stem cell damage, function is killed even death. In addition, stem cell lysery is maintaine...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221
Inventor 高慧晏金红曹秀琴常青马文智何瑞付旭锋俞晓丽陈国栋卢晓华蒲静刘心蕊潘朋歌李雪孙苗狄良娇刘晓萱向袁权菲芸魏爽爽
Owner NINGXIA MEDICAL UNIV