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Breast cancer stem cell cryopreservation protection agent and prepared cryopreservation protection kit

A technology of breast cancer stem cells and cryopreservation agents, applied in the field of stem cells, can solve the problems that it is difficult to effectively maintain the biological characteristics of breast cancer stem cells, and it is not suitable for cryopreservation of breast cancer stem cells, so as to promote commercialization and standardization, Strong tumorigenic ability and damage reduction effect

Inactive Publication Date: 2017-02-15
南京千年健干细胞基因工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Unfortunately, these cryoprotectants for stem cells in the prior art are not well suited for cryopreservation of breast cancer stem cells, and it is difficult to effectively maintain the biological characteristics of breast cancer stem cells before cryopreservation

Method used

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  • Breast cancer stem cell cryopreservation protection agent and prepared cryopreservation protection kit

Examples

Experimental program
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Effect test

Embodiment 1

[0019] Example 1: Effects of different cryoprotectants on the survival rate and tumorigenicity of breast cancer stem cells after cryopreservation and resuscitation

[0020] 1. Experimental materials

[0021] Serum-containing medium for enrichment of MCF-7 breast cancer stem cells: Add 10% fetal bovine serum FBS and double antibody, galantamine and lycorine to 150nmol / L in DMEM / F12 culture medium, and the molar ratio of the two is 1 : 1. Add 10ml penicillin-streptomycin solution (100× double antibody, Beijing Legen Biotechnology Co., Ltd.) to 1L culture medium.

[0022] Serum-free medium for enrichment of MCF-7 breast cancer stem cells: DMEM / F12 culture medium with double antibody (concentration as above), recombinant human epidermal growth factor EGF 20g / L, bovine serum albumin BSA4g / L, insulin 5mg / L , B272% (1L medium plus 20ml 50X B27), galantamine and lycorine total 150nmol / L, the molar ratio of the two is 1:1.

[0023] Serum-free medium for passage of MCF-7 breast cance...

Embodiment 2

[0054] Embodiment 2: cryopreservation protection kit

[0055] A breast cancer stem cell cryopreservation protection kit, used in conjunction with fetal bovine serum FBS for cryopreservation and protection of MCF-7 breast cancer stem cells, including aliquoted serum-free medium lyophilized powder, which consists of the following Method preparation: Add conventional growth factors, galantamine and lycorine to DMEM / F12 culture medium to prepare EGF concentration of 20g / L, BSA concentration of 4g / L, insulin concentration of 5mg / L, concentration of 2%, The galantamine concentration is 75nmol / L, the lycorine concentration is 75nmol / L culture medium, at 37 ℃, 5% CO 2 Stored on a shaker under the conditions for 24h, freeze-dried.

[0056] The kit can also be provided with a subpackaged silica gel desiccant or preservative or antioxidant.

[0057] The cryopreservation agent provided by the invention is designed on the basis of MCF-7 breast cancer stem cell enrichment medium, which ...

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Abstract

The invention discloses a breast cancer stem cell cryopreservation protection agent and a prepared cryopreservation protection kit. The cryopreservation protection agent consists of serum-free medium freeze-dried powder and fetal calf serum FBS, wherein the serum-free medium freeze-dried powder is prepared by the following steps: adding conventional growth factors, galanthamine and lycorine into a DMEM / F12 culture solution to obtain a culture solution of which the EGF concentration is 15-25g / L, the BSA concentration is 3-5g / L, the insulin concentration is 4-6mg / L, the galanthamine concentration is 50-100nmol / L and the lycorine concentration is 50-100nmol / L, wherein the conventional growth factors include recombinant human epidermal growth factor EGF, bovine serum albumin BSA, insulin and B27; storing in a shaking table for 20-28h at 37 DEG C in the presence of 5% CO2; and performing freeze drying. The cryopreservation protection agent disclosed by the invention can effectively reduce the damage of breast cancer stem cells caused by cryopreservation, the recovered cells have a high survival rate and strong tumorigenic ability and are not much different from those before cryopreservation, and thus the invention is remarkably superior to the prior art.

Description

technical field [0001] The invention belongs to the field of stem cells and relates to cryopreservation of tumor stem cells, in particular to a cryopreservation protection agent for breast cancer stem cells and a prepared cryopreservation protection kit. Background technique [0002] On the same day, the applicant applied for "a reagent and kit for enriching breast cancer stem cells from breast cancer cell lines". Breast cancer stem cells, the cell sphere formation cycle is short (from 7 days in the prior art to 3 days and the formation efficiency is significantly improved), CD44 in microsphere cells + CD24 - / low The content of molecularly marked tumor cells and SP cells is high (both increased from less than 10% in the prior art to more than 50%), and can be developed into a kit to improve the efficiency of scientific researchers in enriching breast cancer stem cells. [0003] This application is to solve another problem: how to cryopreserve the enriched breast cancer ste...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221
Inventor 杨超高云雷其他发明人请求不公开姓名
Owner 南京千年健干细胞基因工程有限公司
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