title: [Differentiation of porcine amniotic fluid stem cells into the beating cardiomyocytes].
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.
authors.affiliation: Shaanxi Center for Stem Cell Engineering and Technology, College of Veterinary Medicine, Northwest A&F University, Yangling 712100, China.
authors.forename: Jiahuan
authors.initials: J
authors.lastname: Chen
authors.forename: Yulei
authors.initials: Y
authors.lastname: Wei
authors.forename: Sha
authors.initials: S
authors.lastname: Peng
authors.forename: Huayan
authors.initials: H
authors.lastname: Wang
mesh_terms: D000653:Amniotic Fluid / Q000166:cytology*; D000818:Animals; D001205:Ascorbic Acid / Q000494:pharmacology; D001374:Azacitidine / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects*; D002478:Cells, Cultured; D058732:Embryoid Bodies; D053595:Embryonic Stem Cells / Q000166:cytology*; D005260:Female; D032383:Myocytes, Cardiac / Q000166:cytology*; D013552:Swine
chemical_list: D001374:Azacitidine; D001205:Ascorbic Acid