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Culture medium for preparing endometrial stem cells as well as preparation method

A technology of endometrium and culture medium, applied in cell dissociation methods, cell culture active agents, non-embryonic pluripotent stem cells, etc., can solve the problems of limiting the application prospects of endometrial stem cells and the low survival rate of endometrial stem cells , to achieve the effect of promoting development and application and improving survival rate

Inactive Publication Date: 2019-03-12
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, the survival rate of endometrial stem cells prepared or isolated by the prior art is not high, which limits the application prospect of endometrial stem cells

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1: culture medium of the present invention

[0029] Based on Lonza serum-free medium, containing 2% (w / v) Pall serum replacement, 1% (w / v) penicillin / streptomycin, 1% (w / v) gentamicin sulfate, 2mM gluten Aminoamide, 2 mM MEM NEAA, 1 mM sodium pyruvate, 1 μg / ml EGF, 1 μg / ml bEGF and 1 μg / ml VEGF.

[0030] Among them, Lonza was purchased by the Swiss Lonza Group, batch number: 0000699907; Pall serum substitute was purchased by the American PALL Company, batch number: 259509 / R349-01; penicillin / streptomycin was purchased by Israel BI, batch number: 1807212; gentamicin sulfate Solarbio Cat. No. G8170; Glutamine sigma Cat. No. RNBG6180; MEM NEAA for Gibco Cat. No. 195809; Sodium Pyruvate for Sigma Cat. No. N1633-1

Embodiment 2

[0031] Embodiment 2: The method for preparing endometrial stem cells according to the present invention

[0032] Wash the obtained endometrial tissue material, add 10 times the volume of pre-cooled sodium chloride injection containing 2× double antibody and 0.1% fetal bovine serum, oscillate and shake well, absorb the cleaning solution; then add 10 times the volume of 75% alcohol, soak for no more than 30s, discard the alcohol; finally add tissue cleaning solution and wash twice;

[0033] Cut the washed endometrial tissue to 20-50mm 3, add an equal volume of mixed digestion solution, the digestion solution contains 4mg / ml collagenase I and 2mg / ml Dispase II, digest in a constant temperature shaker at 200R, 37℃ for 30-90min, and obtain a single cell suspension; the digested single cell Pass the cell suspension through a 100 μm disposable filter, discard tiny tissue pieces; centrifuge at 500 g for 10 min, resuspend the cell pellet with Lonza serum-free medium and inoculate it i...

Embodiment 3

[0035] Example 3: Comparative experiment on the survival rate of endometrial stem cells prepared by different techniques

[0036] Experimental group: embodiment 2 method:

[0037] Control group 1: the method is the same as in Example 2, the difference is that α-MEM is the basal medium, 2% Pall serum substitute, 1% (w / v) penicillin / streptomycin, 1% (w / v) cyanine sulfate Amamicin, 2mM glutamine, 2mM MEM NEAA, 1mM sodium pyruvate, 1μg / ml EGF, 1μg / ml PDGF-BB;

[0038] Control group 2: The method is the same as in Example 2, except that the mixed digestive juice contains 4 mg / ml collagenase I and 2 mg / ml collagenase IV;

[0039] Control group 3: the method is the same as in Example 2, the difference is that 4mg / ml collagenase I and 2mg / ml collagenase IV are contained in the mixed digestive juice; Bovine serum, 1% (w / v) penicillin / streptomycin, 1% (w / v) gentamicin sulfate;

[0040] Control group 4: the method is the same as in Example 2, the difference is that 4mg / ml collagenase ...

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PUM

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Abstract

The invention relates to the technical field of tissue culture, and discloses a culture medium for preparing endometrial stem cells as well as a preparation method. On the basis of a serum-free culture medium, Pall serum substitute, penicillin / streptomycin, gentamicin sulfate, glutamine, MEM NEAA, EGF, bEGF, sodium pyruvate and VEGF are added as nutritional ingredients of the endometrial stem cells, so that the survival rate of the endometrial stem cells can be significantly improved; and by combining with the mixed digestion of collagenase I and Dispase II, the survival rate of the endometrial stem cells can be further improved, and the development application of the endometrial stem cells can be promoted.

Description

technical field [0001] The invention relates to the technical field of tissue culture, in particular to a medium for preparing endometrial stem cells and a preparation method. Background technique [0002] Stem cells refer to cells that can self-renew and proliferate indefinitely under certain conditions, including various undifferentiated cells from embryonic development to adult growth and development. The existence, self-renewal and developmental differentiation of stem cells run through the whole life process from fertilized egg to embryonic development until final aging and death. Stem cell research is the fastest growing and most valued cutting-edge biotechnology in the field of life sciences in recent years. It involves all fields of life sciences and biomedicine, and plays an important role in cell therapy, organ transplantation, gene therapy, and new drug screening. [0003] Stem cells can be divided into embryonic stem cells and adult stem cells according to their...

Claims

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Application Information

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IPC IPC(8): C12N5/074
CPCC12N5/0682C12N2500/90C12N2501/11C12N2501/115C12N2501/165C12N2501/999C12N2509/00
Inventor 葛啸虎陈海佳张梦晨王小燕姜交华
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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