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Stem cell cryopreservation method and cell cryopreservation solution used by stem cell cryopreservation method

A cryopreservation method and stem cell technology, applied in the field of stem cell preservation and cultivation, can solve the problems of affecting the effect of clinical treatment, introducing pollution and allergens, and being unsuitable for application, so as to maintain cell osmotic pressure, prolong cryopreservation time, and prevent mechanical damage effect

Inactive Publication Date: 2020-10-30
JIANGSU RE STEM BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, fetal bovine serum is a heterogeneous substance, and there is a risk of introducing contamination and allergens, which may cause unknown adverse reactions and affect the effect of clinical treatment, so it is not suitable for clinical application

Method used

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  • Stem cell cryopreservation method and cell cryopreservation solution used by stem cell cryopreservation method
  • Stem cell cryopreservation method and cell cryopreservation solution used by stem cell cryopreservation method
  • Stem cell cryopreservation method and cell cryopreservation solution used by stem cell cryopreservation method

Examples

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Embodiment

[0042] Example: cryopreservation of stem cells, the cell cryopreservation solution used includes the following components: liquid I and liquid II, wherein liquid I includes human albumin, bFGF and EGF, and the concentration of bFGF is: 1% w / v, the concentration of EGF is: 1% w / v, and the volume is adjusted to 100ml with human serum albumin.

[0043] II solution includes components: Knockout Serum replacement, cytochrome C, betaine, antifreeze protein, glucose, low molecular weight dextran, mannan, sodium chloride, sodium lactate, potassium chloride, calcium chloride and Dimethyl sulfoxide, wherein: the concentration of cytochrome C is: 1.2% w / v, the concentration of betaine is: 0.3% w / v, the concentration of antifreeze protein is: 0.3% w / v, the concentration of glucose is: : 3.5% w / v, low molecular weight dextran concentration: 1.5% w / v, mannan concentration: 1% w / v, sodium chloride concentration: 0.1% w / v, sodium lactate Concentration of: 0.1% w / v, concentration of sodium l...

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Abstract

The invention discloses a stem cell cryopreservation method and a cell cryopreservation solution used by the stem cell cryopreservation method, and belongs to the technical field of stem cell preservation and culture. The stem cell cryopreservation method comprises the following steps: S1, preparing a solution I; S2, preparing a solution II; S3, taking human adipose tissue-derived stromal cells, washing the cells with PBS once, digesting the cells with 0.05% trypsin to change the cells from fusiform to roundness to obtain a cell suspension, and then centrifuging the cell suspension; S4, takingthe solution I to re-suspend the cells; S5, taking the solution II to resuspend the cells; and S6, putting a cryopreservation tube into a programmed cooling box; putting the cell cryopreservation tube in liquid nitrogen for long-term preservation. The cell cryopreservation solution comprises a liquid I and a liquid II, wherein the liquid I comprises human serum albumin, bFGF and EGF, and the liquid II comprises serum replacement cytochrome C, betaine, antifreeze protein, glucose, low-molecular-weight glucan, mannan, sodium chloride, sodium lactate, potassium chloride, calcium chloride and dimethyl sulfoxide. The method has the advantage of improving the cryopreservation quality and stability of the cells.

Description

technical field [0001] The invention relates to the technical field of stem cell preservation and cultivation, in particular to a stem cell cryopreservation method and a cell cryopreservation solution used therefor. Background technique [0002] Stem cells are a type of pluripotent cells with self-replicating ability. Under certain conditions, they can be differentiated into a variety of functional cells. According to the developmental stage of stem cells, they can be divided into embryonic stem cells and adult stem cells. According to the developmental potential of stem cells, they can be divided into totipotent stem cells, pluripotent stem cells and unipotent stem cells. [0003] Adipose-derived mesenchymal stem cells are adult pluripotent stem cells that exist in adipose tissue, and can differentiate into bone, cartilage, cardiac muscle, nerve, fat and other tissue cells under certain conditions. At present, adipose-derived mesenchymal stem cells have made a lot of prog...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221A01N1/0226
Inventor 焦阳王铁朱迪·德瓦库玛杨丽华李靖
Owner JIANGSU RE STEM BIOTECH
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