Serum-free induced pluripotent stem cell cryopreservation liquid and cryopreservation method

A technology for pluripotent stem cells and cryopreservation, applied in the field of induced pluripotent stem cell cryopreservation, can solve the problems of residual pluripotent stem cells, protein denaturation, cell damage, etc., to maintain cell viability, prevent cell differentiation, and reduce cell proliferation. damage effect

Inactive Publication Date: 2018-09-21
GUANGDONG XTEM BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, high concentrations of dimethyl sulfoxide are toxic and can interact with the hydrophobic groups of intracellular proteins, leading to protein denaturation, cell damage or inactivation
Moreover, the use of animal-derived serum in the existing cryopreservation solution leads to the addition of non-personnel substances into the cell cryopreservation solution, which will cause residue or contamination of pluripotent stem cells

Method used

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  • Serum-free induced pluripotent stem cell cryopreservation liquid and cryopreservation method
  • Serum-free induced pluripotent stem cell cryopreservation liquid and cryopreservation method
  • Serum-free induced pluripotent stem cell cryopreservation liquid and cryopreservation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1. Preparation of Induced Pluripotent Stem Cell Cryopreservation Solution

[0032] The freezing solution consists of liquid A and liquid B. The volume ratio of liquid A to liquid B is 1:1.

[0033] Each 100mL Freezing Solution A contains the following components:

[0034]

[0035] Each 100mL freezing solution B contains the following components:

[0036]

[0037]

[0038] Preparation method: Mix all ingredients to obtain liquid A and liquid B respectively, and store at 4°C.

Embodiment 2

[0039] Example 2. Application of cryopreservation solution in cryopreservation of induced pluripotent stem cells

[0040] Freezing method:

[0041] After digesting the induced pluripotent stem cells in the logarithmic growth phase, centrifuge at 800rpm / min for 5 minutes to remove the supernatant, resuspend with an appropriate amount of liquid A of the present invention, and perform cell counting, then add an equal amount of liquid B to make the cells The concentration is 1×10 6 -6×10 6 cells / mL, and then 1 mL / tube was dispensed into 1.8 mL cell cryopreservation tubes, placed in a programmed cooling box at -80°C for programmed cooling, and transferred to liquid nitrogen for storage after 24 hours.

[0042] Inspection of frozen storage results:

[0043] 1. Morphological observation of pluripotent stem cells after resuscitation

[0044] The frozen cells were resuscitated, and the medium was changed every day, and the morphology was observed under a microscope, and pictures we...

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PUM

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Abstract

The invention provides an induced pluripotent stem cell cryopreservation liquid and a cryopreservation method thereof. The cryopreservation liquid is composed of a DMEM basal culture medium, NEAA, KSR, DMSO, hydroxyethyl starch and the like, is clear and definite in ingredients and free of contamination by animal serum, and has very high application value. The cryopreservation method is easy to operate, repeated freezing and thawing are not needed, and the cryopreservation liquid can be utilized right after it is ready. Cells obtained through cryopreservation of the cryopreservation liquid andthe cryopreservation method have the advantages that the cell viability is good and the proliferation speed is high. The induced pluripotent stem cell cryopreservation liquid and the cryopreservationmethod thereof are significant in prompting the study on induced pluripotent stem cells and building a bank of the induced pluripotent stem cells.

Description

technical field [0001] The invention relates to the fields of stem cell biology and regenerative medicine, in particular to a cryopreservation liquid for induced pluripotent stem cells and a cryopreservation method for induced pluripotent stem cells. Background technique [0002] Induced pluripotent stem cells refer to pluripotent stem cells obtained by reprogramming differentiated somatic cells, which have the potential of self-renewal and multi-directional differentiation. In many ways, they are similar to embryonic stem cells in terms of their ability to differentiate and multilineage. Induced pluripotent stem cells are not restricted by many aspects such as cell source, immune rejection, ethics, religion and law. They have great application value in drug development, disease treatment and mechanism research, and are currently a hot spot in stem cell research worldwide. [0003] Since the acquisition of pluripotent stem cells is time- and labor-intensive, and the cell ph...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/021
Inventor 刘樱彭特陈勇蔡亚雄于云飞乔志平
Owner GUANGDONG XTEM BIOTECH CO LTD
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