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Human induced multipotential stem cells cryopreserved reagent and cryopreserving method thereof

A technology of pluripotent stem cells and cryopreservation method, which is applied in the field of cryopreservation reagents and cell cryopreservation, can solve the problems of unstable cell state, unstable batch, cumbersome operation, etc., and achieve simple and easy-to-operate cryopreservation method and convenient quality Effects of controlling and preventing cell differentiation

Inactive Publication Date: 2017-06-30
北京焕生汇生物技术研究院有限公司
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Problems solved by technology

[0004] At present, 90% serum plus 10% DMSO is generally used as the cryopreservation solution for cryopreservation of human induced pluripotent stem cells. Cell status is unstable during subsequent cell recovery

Method used

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  • Human induced multipotential stem cells cryopreserved reagent and cryopreserving method thereof
  • Human induced multipotential stem cells cryopreserved reagent and cryopreserving method thereof
  • Human induced multipotential stem cells cryopreserved reagent and cryopreserving method thereof

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Embodiment Construction

[0032] The present invention will be specifically introduced below in conjunction with the accompanying drawings and specific embodiments.

[0033] 1. Preparation of frozen storage reagents

[0034] Each 100ml frozen reagent contains the following components:

[0035]

[0036] The composition of the whole formula is completely clear, which is convenient for quality control, avoids containing animal serum components, is safe and reliable, and is suitable for clinical application.

[0037] Preparation method: take DMEM-F12 medium, add trehalose, recombinant human albumin, recombinant human epidermal growth protein, b-mercaptoethanol and dimethyl sulfoxide in sequence, and mix evenly.

[0038] In the formula of the present invention:

[0039] Recombinant human albumin replaces the fetal bovine serum of animal components, avoids the existence of animal components, is safer, and at the same time allows cells to obtain sufficient nutrients;

[0040] Recombinant human epidermal...

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Abstract

The invention discloses a human induced multipotential stem cells cryopreserved reagent and a cryopreserving method thereof. The cryopreserved reagent comprises the following components: 0.03-100 ml of recombinant human albumin, 0.0125 mg / 100 ml of recombinant human epidermal growth protein, 1 ml / 100 ml of beta-mercaptoethanol, 91-100 ml of a DMEM-F12 medium, 8-100 ml of dimethyl sulfoxide, and 5-100 ml of trehalose. The method has the beneficial effects that 1) the component is completely clear and is convenient for quality control; 2) the recombinant human albumin is used for substituting the fetal calf serum of the animal component, can avoid an animal serum-containing component, the cryopreserved reagent is safe and reliable, and is adapted to clinic application; 3) the recombinant human epidermal growth protein, beta-mercaptoethanol, and trehalose in a formula can better protect the cells in a state of dormancy under low temperature environment, ice crystal is not generated in tissue of the cells, cell activity of the cells can be kept, the cell differentiation can be effectively prevented, and the recovery rate of the cryopreserved cell can reach more than 96%; and 4) the cryopreserved reagent belongs to a ready-to-use type, and the usage is convenient.

Description

technical field [0001] The invention relates to a cryopreservation reagent and a cell cryopreservation method, in particular to a special cryopreservation reagent for human induced pluripotent stem cells, and a method for using the cryopreservation reagent to freeze human induced pluripotent stem cells, belonging to cell biology field of technology. 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 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 pluripotent stem cells, cell cryopreservation is often involved. The process of cell cryopreservation will significantly change the thermodynamic, chemical and physical environment of c...

Claims

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Application Information

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IPC IPC(8): A01N1/02
CPCA01N1/0221A01N1/0226
Inventor 姚玲玲杨敏孟得龙杨苗
Owner 北京焕生汇生物技术研究院有限公司
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