Cell culture solution and application thereof, and method for inducing differentiation of skeletal muscle stem cells into cardiomyocyte-like cells

A cardiomyocyte-like cell and cell culture technology, applied in the field of stem cell culture, can solve the problem of low transformation efficiency of skeletal muscle stem cells

Inactive Publication Date: 2016-12-21
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the current culture method, the conversion efficiency of skeletal muscle stem cells to cardiomyocyte-like cells is still very lo...

Method used

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  • Cell culture solution and application thereof, and method for inducing differentiation of skeletal muscle stem cells into cardiomyocyte-like cells
  • Cell culture solution and application thereof, and method for inducing differentiation of skeletal muscle stem cells into cardiomyocyte-like cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Isolation and passage of embodiment 1MDSCs

[0041] 1. Materials:

[0042] Take adult normal temporal muscle specimens (taken from craniotomy patients), rinse with PBS buffer containing double antibodies, transfer them to sterile petri dishes, and cut them into 1mm with ophthalmic scissors 3 Fragments of about the size are then transferred to a 50ml centrifuge tube, washed with PBS buffer for 3 times, and left to stand for 1 minute before discarding the supernatant and floating tissues.

[0043] 2. Enzyme hydrolysis:

[0044] Add 2 times the volume of mixed enzymes to the muscle fragments obtained above, including 2.4u / ml Dispase II, 1% type I collagenase, 2.5mmol / L CaCl2, mix well and place in a constant temperature shaker at 37°C for about 60 minutes to digest , until the muscle fragments in the test tube are digested into minced muscle, and the muscle mass cannot be seen with the naked eye. After digestion, centrifuge at 1000r / min for 5min and discard the supernat...

Embodiment 2

[0052] The differentiation of embodiment 2MDSCs

[0053] The formula of the culture medium of each test group is shown in Table 1:

[0054] Table 1 The formulation of each component differentiation induction solution

[0055] group

basal medium

FBS

5-aza

TGFβ

Experimental group 1

DMEM / F12

10％

5umol / L

1ug / L

Experimental group 2

DMEM / F12

10％

10umol / L

5ug / L

Experimental group 3

DMEM / F12

10％

5umol / L

10ug / L

Experimental group 4

DMEM / F12

10％

15umol / L

10ug / L

Control group 1

DMEM / F12

10％

——

——

Control group 2

DMEM / F12

10％

10umol / L

——

Control group 3

DMEM / F12

10％

——

5ug / L

[0056] In order to verify the effect of differentiation induction of the present invention, 3 groups of control groups and 4 groups of experimental groups were set up at the same time, and the third generation MDSCs were selected, and the ...

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Abstract

The invention relates to the technical field of stem cell culture, particularly a method for inducing differentiation of skeletal muscle stem cells into cardiomyocyte-like cells. The culture solution comprises a base culture solution, FBS (fetal bovine serum), 5-aza and a TGF (transforming growth factor) beta. The culture solution can induce differentiation of skeletal muscle stem cells into cardiomyocyte-like cells, and is capable of enhancing the differentiation rate from skeletal muscle stem cells to cardiomyocyte-like cells and shortening the differentiation time. After induction for four weeks, the differentiation rates of every group of cells are calculated, and the result shows that the differentiation rate of the cells in the experimental group is obviously higher than the control group (p<0.05 or p<0.01).

Description

technical field [0001] The invention relates to the technical field of stem cell culture, in particular to a cell culture solution and its application and a method for inducing skeletal muscle stem cells to differentiate into cardiomyocyte-like cells. Background technique [0002] Myocardial infarction is the myocardial damage caused by the imbalance between coronary blood flow and myocardial demand caused by changes in coronary circulation. It is a serious clinical ischemic heart disease. Cardiomyocytes after necrosis are gradually replaced by scar tissue. Due to the lack of elasticity of scar tissue, it is difficult to meet the requirements of cardiac contraction and relaxation. In order to compensate for the loss of cardiomyocyte function, the heart gradually undergoes degenerative left ventricular remodeling. decline, eventually leading to congestive heart failure and even sudden death. Although clinical drugs and interventional therapy can improve symptoms, they cannot...

Claims

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Application Information

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IPC IPC(8): C12N5/077C12N5/0775
CPCC12N5/0657C12N2501/06C12N2501/15C12N2506/1392
Inventor 葛啸虎陈海佳王一飞戚康艺
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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