Special thawing reagent and thawing method for human induced pluripotent stem cells

A pluripotent stem cell and inducible technology, applied in the field of cell biology, can solve the problems of resuscitation survival rate (low recovery rate, no resuscitation reagent, poor versatility, etc.), achieve good versatility, widely popularize and use, and high survival rate Effect

Active Publication Date: 2017-08-11
焕生汇生物基因技术(北京)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] (1) A resuscitation reagent is only suitable for one type of cell, or only for a few similar cells, and the versatility is poor;
[0011] (2) After cell recovery, the recovery survival rate (recovery rate) is low, and the activity is poor
[0012] For human induced pluripotent stem cells, there is currently no specific resuscitation reagent, and other cell resuscitation reagents used in resuscitation procedures also contain animal components, which are not safe to use

Method used

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  • Special thawing reagent and thawing method for human induced pluripotent stem cells
  • Special thawing reagent and thawing method for human induced pluripotent stem cells
  • Special thawing reagent and thawing method for human induced pluripotent stem cells

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Experimental program
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Effect test

Embodiment 1

[0051] Accurately weigh 100ng of recombinant human epidermal growth factor and 1000ng of recombinant human serum albumin, first add 5ml of DMEM-F12, mix gently, add 0.1ml of b-mercaptoethanol after complete dissolution, and finally supplement with DMEM-F12 medium 10.0ml, mix thoroughly.

Embodiment 2

[0053] Accurately weigh 200ng of recombinant human epidermal growth factor and 1500ng of recombinant human serum albumin, first add 5ml of DMEM-F12, mix gently, add 0.2ml of b-mercaptoethanol after complete dissolution, and finally supplement with DMEM-F12 medium 10.0ml, mix thoroughly.

Embodiment 3

[0055] Accurately weigh 300ng of recombinant human epidermal growth factor and 2000ng of recombinant human serum albumin, first add 5ml DMEM-F12 and mix gently, then add 0.25ml of b-mercaptoethanol after completely dissolving, and finally supplement 10.0ml with DMEM-F12 medium ml, and mix well.

[0056] In the formula of the special resuscitation reagent for human induced pluripotent stem cells of the present invention:

[0057] (1) The role of recombinant human epidermal growth factor and b-mercaptoethanol is to protect the cells from dormant state in low temperature environment, the internal structure of the cells does not produce ice crystals, so as to maintain the activity of the cells and ensure that the cells still maintain good multi-functionality sex.

[0058] (2) The function of recombinant human serum albumin is to replace the fetal bovine serum of animal components, avoid the existence of animal components, be safer, and at the same time allow cells to obtain suffi...

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Abstract

The invention discloses a special thawing reagent and a thawing method for human induced pluripotent stem cells. The thawing reagent is free of animal components and comprises 100ng-300ng of recombinant human epidermal growth factors, 1000ng-2000ng of recombinant human blood albumin, 0.1ml-0.25ml of b-mercaptoethanol and the balance DMEM-F12 culture medium, wherein the total volume is 10.0ml. The special thawing reagent and the thawing method for the human induced pluripotent stem cells have advantages that (1) high survival rate (thawing rate) and high activity of the human induced pluripotent stem cells can be guaranteed, and high functional versatility of the cells is guaranteed as well; (2) due to freeness of the animal components, complete exactness and controllability of components can be realized, and high safety in use is achieved; (3) the thawing method is convenient to operate, easy to implement and suitable for wide popularization and utilization.

Description

technical field [0001] The invention relates to a cell resuscitation reagent and a resuscitation method, in particular to a special resuscitation reagent for human induced pluripotent stem cells and a method for resuscitating human induced pluripotent stem cells by using the resuscitation reagent, belonging to the technical field of cell biology. Background technique [0002] Human induced pluripotent stem cells are the hotspot and focus of current stem cell research. It can differentiate into all cells in the body, and then form all tissues and organs of the body. Therefore, the study of human induced pluripotent stem cells not only has important theoretical significance, but also has great application value in organ regeneration, repair and disease treatment. [0003] During the research process of human induced pluripotent stem cells, cell cryopreservation and recovery of cryopreserved cells are often involved. [0004] 1. Cell cryopreservation [0005] There are two t...

Claims

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Application Information

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IPC IPC(8): C12N5/10
CPCC12N5/0696C12N2500/44C12N2501/11C12N2501/998C12N2510/00
Inventor 姚玲玲杨敏孟得龙杨苗
Owner 焕生汇生物基因技术(北京)有限公司
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