title: [Involvement of human amniotic fluid colony derived stem cells in regeneration of mouse injured muscle].
abstract: OBJECTIVE
  To study whether human amniotic fluid colony derived stem cells (hAFCSCs) are involved in regeneration of injured muscles in mice and to investigate the method and feasibility of hAFCSCs-based cytotherapy in the treatment of injured muscles.
  
  
  METHODS
  s Human second-trimester amniotic fluid was collected through ultrasound-guided amniocentesis, hAFCSCs were isolated from second-trimester amniotic fluid and cultured, and the cells at 6th-8th passages were spared. The mRNA was extracted to identify the stem cell related genes by RT-PCR. The muscular injury model of bilateral tibialis anterior muscle was established by cardiotoxin and X-ray irradiation in 16 Nod/Scid mice (aged 6-8 weeks, and weighing 20-24 g). The hAFCSCs (3.3 x 1(0)7/mL, 3 microIL) were injected into the right injured tibialis anterior muscles as the experimental group, while the same volume of complete medium (lphaa-MEM containing 15%FBS, 18%Chang B, 2%Chang C, 1% penicillin-streptomycin, and 1% L-glutamine) was injected into the left injured tibialis anterior muscles as the control group. At 2 and 4 weeks after cell transplantation, the immunofluorescence staining of tibialis anterior muscles was performed to detect hepatocyte growth factor receptor (c-Met), myogenic regulatory factor (Myf-5), Laminin, Desmin, and human specific nuclear mitotic apparatus protein (NuMa).
  
  
  RESULTS
  s The clone formation was observed at 5-7 days of primary hAFCSCs culture; after 8-10 days, the clones with homogeneous morphology were selected for subculture. Adequate stem cells were available after 6th-8th subculture. RT-PCR analysis showed that hAFCSCs expressed mRNA of the stem cell related genes. The immunofluorescence double-staining showed that NuMa expressed in tibialis anterior muscles of the experimental group and no myogenic phenotype expressed at 2 weeks after cell transplantation, and that single cell co-expressed NuMa and c-Met or Myf-5 at 4 weeks after cell transplantation. In some myofibers, NuMa and Laminin or Desmin were also co-expressed. No NuMa positive hAFCSCs were detected in the control group at 2 and 4 weeks after cell transplantation.
  
  
  CONCLUSIONS
  n hAFCSCs can participate in the regeneration of injured mouse muscle.
authors.affiliation: Central Laboratory, Shanghai Children's Hospital Affiliated to Shanghai Jiaotong University, Shanghai, 200040, P.R.China.
authors.forename: Xiaorong
authors.initials: X
authors.lastname: Ma
authors.forename: Shengli
authors.initials: S
authors.lastname: Zhang
authors.forename: Yafeng
authors.initials: Y
authors.lastname: Shang
authors.forename: Tongbin
authors.initials: T
authors.lastname: Gao
authors.forename: Xue
authors.initials: X
authors.lastname: Wang
authors.forename: Fang
authors.initials: F
authors.lastname: Chen
authors.forename: Junmei
authors.initials: J
authors.lastname: Zhou
mesh_terms: D000653:Amniotic Fluid / Q000166:cytology*; D000818:Animals; D017690:Cell Transplantation; D002478:Cells, Cultured; D006801:Humans; D051379:Mice; D016513:Mice, SCID; D018482:Muscle, Skeletal / Q000293:injuries* / Q000502:physiology*; D012038:Regeneration; D013234:Stem Cells / Q000166:cytology*