Freezing medium for induced pluripotent stem cells, application of freezing medium, and freezing method of induced pluripotent stem cells

A technology of pluripotent stem cells and cryopreservation method, which is applied in the field of induced pluripotent stem cell cryopreservation solution, to achieve high viability, high use value, and the effect of avoiding the risk of animal-derived pathogens

Active Publication Date: 2015-11-25
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, fetal bovine serum is derived from animals and may contain some animal

Method used

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  • Freezing medium for induced pluripotent stem cells, application of freezing medium, and freezing method of induced pluripotent stem cells
  • Freezing medium for induced pluripotent stem cells, application of freezing medium, and freezing method of induced pluripotent stem cells
  • Freezing medium for induced pluripotent stem cells, application of freezing medium, and freezing method of induced pluripotent stem cells

Examples

Experimental program
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Effect test

Embodiment 1

[0043] The raw material of dextran 40 is 6% dextran 40-sodium chloride injection (Guosen Pharmaceutical).

[0044] 10ug Thiazovivin (StemRD Company, USA) was dissolved in 1ml DMSO to make 1000 times Thiazovivin storage solution.

[0045] Add 16.67ml dextran 40 injection, 5ml albumin injection, 100ul Thiazovivin storage solution to 68.23ml DMEM / F12 basal medium, and mix well; then slowly add 10ml DMSO solution, mix well, and store in a 4 degree refrigerator for later use.

Embodiment 2

[0047] The raw material of dextran 40 is 6% dextran 40-sodium chloride injection (Guosen Pharmaceutical).

[0048] 10ug Thiazovivin (StemRD Company, USA) was dissolved in 1ml DMSO to make 1000 times Thiazovivin storage solution.

[0049] Add 8.335ml dextran 40 injection, 2.5ml albumin injection, 10ul Thiazovivin storage solution to 69.155ml DMEM / F12 basal medium, and mix well; then slowly add 20ml DMSO solution, mix well, and store in a 4 degree refrigerator for later use.

Embodiment 3

[0051] The raw material of dextran 40 is 6% dextran 40-sodium chloride injection (Guosen Pharmaceutical).

[0052] 10ug Thiazovivin (StemRD Company, USA) was dissolved in 1ml DMSO to make 1000 times Thiazovivin storage solution.

[0053] Add 33.34ml dextran 40 injection, 25ml albumin injection, 10ml Thiazovivin storage solution to 29.66ml DMEM / F12 basic medium, and mix well; then slowly add 2ml DMSO solution, mix well, and store in a 4 degree refrigerator for later use.

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Abstract

The invention relates to the field of biology and provides a freezing medium for induced pluripotent stem cells, application of the freezing medium, and a freezing method of the induced pluripotent stem cells. The freezing medium uses an IMDM/F12 basal medium as substrate; each 100mL of the freezing medium comprises, by volume concentration, 2-20v/v% of DMSO, 0.5-5v/v% of dextran 40, 0.5-5v/v% of albumin, 1ng-1000ng of Thiazovivin, and the balance of IMDM/F12 basal medium. The freezing medium uses no animal serum, thus the risk of serum spreading of animal-origin pathogens is avoided. In addition, freezing with the freezing medium is more effective, and post-thawing survival rate of the cells is higher. After three months of freezing, the survival rate of thawed iPS (induced pluripotent stem) cells frozen with the freezing medium is higher than 91%, which is evidently higher than that of traditional freezing medium.

Description

Technical field [0001] The present invention relates to the field of biotechnology, in particular to a cryopreservation solution for induced pluripotent stem cells, its application and a cryopreservation method. Background technique [0002] Induced pluripotent stem cells (iPScells) were originally the Japanese Shinya Yamanaka (Shinya Yamanaka) used viral vectors to transfer a combination of four transcription factors (Oct4, Sox2, Klf4 and c-Myc) into differentiated somatic cells in 2006 , A cell type similar to embryonic stem cells obtained by reprogramming. Induced pluripotent stem cells can reprogram somatic cells to obtain pluripotent stem cells with embryonic stem cell-like characteristics by introducing inducible genes into somatic cells, also known as dedifferentiation. [0003] Subsequently, different scientists around the world have discovered that other methods can also be used to make such cells. After successful establishment of iPS cells, the use of iPS cells to trea...

Claims

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Application Information

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IPC IPC(8): C12N5/0735
Inventor 王一飞陈海佳葛啸虎卢瑞珊王小燕李平
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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