Differentiation of porcine amniotic fluid stem cells into the beating cardiomyocytes.
- Author:
Jiahuan CHEN
1
;
Yulei WEI
;
Sha PENG
;
Huayan WANG
Author Information
1. Shaanxi Center for Stem Cell Engineering and Technology, College of Veterinary Medicine, Northwest A&F University, Yangling 712100, China.
- Publication Type:Journal Article
- MeSH:
Amniotic Fluid;
cytology;
Animals;
Ascorbic Acid;
pharmacology;
Azacitidine;
pharmacology;
Cell Differentiation;
drug effects;
Cells, Cultured;
Embryoid Bodies;
Embryonic Stem Cells;
cytology;
Female;
Myocytes, Cardiac;
cytology;
Swine
- From:
Chinese Journal of Biotechnology
2011;27(8):1206-1214
- CountryChina
- Language:Chinese
-
Abstract:
The aim of this research is to find an effective cardiomyocyte-induced method derived from porcine amniotic fluid stem cells (pAFS). For cardiac differentiation, the cells were formed embryoid bodies (EBs) firstly, then cultured in induced-medium including 5-azacytidine (5-aza) and vitamin C (Vc). We detected the specific markers of cardiomyocyte by immunocytochemistry, RT-PCR and transmission electron microscope. The results showed that some embryoid bodies beat rhythmically after 10 days of induction. Furthermore, analysis of t test revealed that the percentage of beating cardiomyocyte-like cell clusters was highest (33%) when induction using 0.1 mmol/L Vc and 5 micromol/L 5-aza. Immunocytochemistry analysis demonstrated that cardiomyocyte-like cell clusters expressed alpha-actin, Tnni3. RT-PCR analysis also illustrated that TbX5, Gata4, alpha-MHC and Tnni3 were expressed positive in cardiomyocyte-like cell clusters. Especially, we observed basic structures of myocardium, such as myofilament, glycogen granule and so on by transmission electron microscope. In conclusion, 5-azacytidine and vitamin C could promote differentiation of pAFS into myocardium.
