title: [Cardio-myogenic differentiation of human amniotic fluid colony derived stem cells in the form of embryonic body-like structure].
abstract: OBJECTIVE
  To investigate cardio-myogenic differentiation potential of human amniotic fluid colony derived stem cells (HAFCDSC) in the form of embryonic body (EB)-like structure in vitro.
  
  
  METHODS
  HAFCDSC were isolated from second trimester amniotic fluid which was backup of amniocentesis specimens. The forth passage of HAFCDSC were cultured by hanging-drop preparation in complete medium for 5 days to form EB-like structures followed by inducing medium in regular tissue culture dishes for 2 weeks (experiment group). The EB-like structures cultured in complete medium were served as control group. Cardiomyocyte phenotypes were detected by RT-PCR and immunofluorescence staining.
  
  
  RESULTS
  HAFCDSC could form EB-like structures 24 h after hanging drop culture, and the diameter of EB-like structures gradually increased with culture duration and the mean diameter of EB-like structures reached 237.3 ± 26.7 µm on 5(th) day. The formation rate of EB-like structures was 93.5%. RT-PCR analysis showed EB-like structures expressed stem cell related mRNA. Immunofluorescence staining demonstrated that some cardiomyocyte phenotypes such as Desmin, α-Actinin and Tn I were expressed by HAFCDSCs in EB-like structures after cardio-myogenic induction. Cardiomyocytes specific mRNA including Desmin, α-Actin, Tn I, Tn T, T-box and NK2.5 were also transcripted as detected by RT-PCR. No positive results was found in control group.
  
  
  CONCLUSIONS
  HAFCDSC can form uniform EB-like structures after hanging drop culture for 5 days. HAFCDSC differentiate into cardiomyocyte-like cells through the form of EB-like structures after induction by 5-Aza.
authors.affiliation: Shanghai Jiaotong University, Central Laboratory, Shanghai, China.
authors.forename: Xiao-rong
authors.initials: XR
authors.lastname: Ma
authors.forename: Hua
authors.initials: H
authors.lastname: Xie
authors.forename: Sheng-li
authors.initials: SL
authors.lastname: Zhang
authors.forename: Tong-bin
authors.initials: TB
authors.lastname: Gao
authors.forename: Xue
authors.initials: X
authors.lastname: Wang
authors.forename: Ya-feng
authors.initials: YF
authors.lastname: Shang
authors.forename: Jun-mei
authors.initials: JM
authors.lastname: Zhou
authors.forename: Fang
authors.initials: F
authors.lastname: Chen
mesh_terms: D000653:Amniotic Fluid / Q000166:cytology*; D001374:Azacitidine / Q000494:pharmacology; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D058732:Embryoid Bodies; D053595:Embryonic Stem Cells / Q000166:cytology*; D006801:Humans; D032383:Myocytes, Cardiac / Q000166:cytology*
chemical_list: D001374:Azacitidine