title: Stemness and angiogenic gene expression changes of serial-passage human amnion mesenchymal cells.
abstract: BACKGROUND
  Particular attention has been directed towards human amnion mesenchymal stem cells (HAMCs) due to their accessibility, availability and immunomodulatory properties. Therefore, the aim of the present study was to determine the temporal changes of stemness and angiogenic gene expressions of serial-passage HAMCs.
  
  
  METHODS
  HAMCs were isolated from human term placenta and cultured in serial passages in culture medium supplemented with 10% fetal bovine serum. Morphological analysis, growth kinetic and CFU-F assay of HAMCs were assessed. In vitro differentiation and the immunophenotype of HAMCs at P5 were also analyzed. Quantitative PCR was used to determine the stemness, angiogenic and endothelial gene expression of cultured HAMCs after serial passage.
  
  
  RESULTS
  Cultured HAMCs displayed intermediate epitheloid-fibroblastoid morphology at an initial culture and the fibroblastoid features became more pronounced in later passages. They showed high clonogenic activity and faster proliferation at later passages with colony forming efficiency of 0.88%. HAMCs were successfully differentiated into adipocytes, osteocytes and neuron-like cells. Most HAMCs expressed CD9, CD44, CD73, CD90 and HLA-A,B,C but negligibly expressed CD31, CD34, CD45, CD117 and HLA-DR,DP,DQ. After serial passage, stemness genes Oct-3/4, Sox-2, Nanog3, Rex-1, FGF-4 and FZD-9 expressions significantly decreased. Of the angiogenic genes PECAM-1, bFGF, eNOS, VEGFR-2, VEGF, and vWF expressions also decreased significantly except angiopoietin-1 which significantly increased. No significant differences were observed in ABCG-2, BST-1, nestin, PGF and HGF expressions after serial passage.
  
  
  CONCLUSIONS
  These results suggested that cultured HAMCs could be an alternative source of stem cells and may have the potential for angiogenesis and hence its use in stem-cell based therapy.
authors.affiliation: Department of Pathology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Jalan Yaacob Latif, Bandar Tun Razak, Cheras, 56000 Kuala Lumpur, Malaysia. huntress_295@hotmail.com
authors.forename: Simat Siti
authors.initials: SS
authors.lastname: Fatimah
authors.forename: Geok Chin
authors.initials: GC
authors.lastname: Tan
authors.forename: Kienhui
authors.initials: K
authors.lastname: Chua
authors.forename: Mohd Manzor Nur
authors.initials: MM
authors.lastname: Fariha
authors.forename: Ay Eeng
authors.initials: AE
authors.lastname: Tan
authors.forename: Abdul Rahman
authors.initials: AR
authors.lastname: Hayati
mesh_terms: D017667:Adipocytes / Q000166:cytology; D000328:Adult; D000650:Amnion / Q000166:cytology*; D042501:Angiogenic Proteins / Q000096:biosynthesis / Q000235:genetics; D000943:Antigens, Differentiation / Q000096:biosynthesis / Q000235:genetics; D002454:Cell Differentiation / Q000187:drug effects; D002455:Cell Division; D002478:Cells, Cultured / Q000187:drug effects / Q000378:metabolism; D003470:Culture Media / Q000494:pharmacology; D004847:Epithelial Cells / Q000166:cytology; D005260:Female; D005347:Fibroblasts / Q000166:cytology; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D006801:Humans; D016130:Immunophenotyping; D059630:Mesenchymal Stem Cells / Q000187:drug effects / Q000378:metabolism*; D008648:Mesoderm / Q000166:cytology*; D018919:Neovascularization, Physiologic / Q000235:genetics*; D054258:Neural Plate / Q000166:cytology; D009474:Neurons / Q000166:cytology; D010011:Osteocytes / Q000166:cytology; D011247:Pregnancy; D061251:Primary Cell Culture; D055815:Young Adult
chemical_list: D042501:Angiogenic Proteins; D000943:Antigens, Differentiation; D003470:Culture Media