5-Azacytidine combined with angiotensin II induces differentiation of  adipose-derived mesenchymal stem cells into cardiomyocytes

doi:10.3969/j.issn.2095-4344.2017.29.007

Abstract

Abstract:

BACKGROUND: Taking effective measures for myocardial necrosis develops a trend in the clinical treatment of myocardial infarction.
OBJECTIVE: To investigate the mechanism of 5-azacytidine combined with angiotensin II in the differentiation of mouse adipose-derived mesenchymal stem cells into cardiomyocytes.
METHODS: Twenty ICR mice aged 4 weeks old were used to isolate adipose-derived mesenchymal stem cells using type I collagenase digestion method. Cell morphology was observed, cell proliferation was detected using MTT, and immunofluorescence technique was used to detect the expression of CD90, CD105, CD73 and CD45. Passage 4 adipose-derived mesenchymal stem cells were cultured for 24 hours, then induced by 15 μmol/L 5-azacytidine or
15 μmol/L 5-azacytidine plus 0.1 μmol/L angiotensin II for another 24 hours, and finally cultured in a complete medium. The expression of cTnT protein in myocardial cells was detected by western blot assay.
RESULTS AND CONCLUSION: Primary adipose-derived mesenchymal stem cells with small nuclei were spindle-shaped or starlike. After passage, the cells were increased in size, with uniform shape and regular arrangement. Passage 3 mouse adipose-derived mesenchymal stem cells showed an S-shaped growth curve: days 1-3 was incubation period, the cells showed logarithmic growth at day 4, the cell growth reached the peak at days 5-7, and then entered stationary phase. The passage 4 cells were positive for CD90, CD105 and CD73, but negative for CD45. The expression of cTnT was significantly higher after cell induction with 5-azacytidine combined with angiotensin II than 5-azacytidine alone. In conclusion, the combined use of 5-azacytidine and angiotensin II is beneficial for the cardiomyocyte-like differentiation of adipose-derived mesenchymal stem cells.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Adipose Tissue, Mesenchymal Stem Cells, Angiotensin II, Myocytes, Cardiac, Tissue Engineering

CLC Number:

R394.2

Wang Jing, Han Jiang-hong, Li Qiong. 5-Azacytidine combined with angiotensin II induces differentiation of adipose-derived mesenchymal stem cells into cardiomyocytes[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(29): 4629-4634.

Figures/Tables 4

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