Stemness and angiogenic gene expression changes of serial-passage human amnion mesenchymal cells

Simat Siti Fatimah, Geok Chin Tan, Chua Kien Hui, Mohd Manzor Nur Fariha, Ay Eeng Tan, Abdul Rahman Hayati

Research output: Contribution to journalArticle

30 Citations (Scopus)

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. Conclusion: 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.

Original languageEnglish
Pages (from-to)21-29
Number of pages9
JournalMicrovascular Research
Volume86
Issue number1
DOIs
Publication statusPublished - Mar 2013

Fingerprint

Serial Passage
Amnion
Stem cells
Gene expression
Mesenchymal Stromal Cells
Gene Expression
Stem Cells
HLA-DP Antigens
Angiopoietin-1
Genes
CD31 Antigens
Fibroblast Growth Factor 1
Osteocytes
Nestin
Vascular Endothelial Growth Factor Receptor-2
HLA-A Antigens
HLA-B Antigens
Prostaglandins F
HLA-DR Antigens
Cell- and Tissue-Based Therapy

ASJC Scopus subject areas

  • Biochemistry
  • Cardiology and Cardiovascular Medicine
  • Cell Biology

Cite this

Stemness and angiogenic gene expression changes of serial-passage human amnion mesenchymal cells. / Fatimah, Simat Siti; Tan, Geok Chin; Kien Hui, Chua; Fariha, Mohd Manzor Nur; Tan, Ay Eeng; Hayati, Abdul Rahman.

In: Microvascular Research, Vol. 86, No. 1, 03.2013, p. 21-29.

Research output: Contribution to journalArticle

Fatimah, Simat Siti ; Tan, Geok Chin ; Kien Hui, Chua ; Fariha, Mohd Manzor Nur ; Tan, Ay Eeng ; Hayati, Abdul Rahman. / Stemness and angiogenic gene expression changes of serial-passage human amnion mesenchymal cells. In: Microvascular Research. 2013 ; Vol. 86, No. 1. pp. 21-29.
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AU - Tan, Geok Chin

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AU - Tan, Ay Eeng

AU - Hayati, Abdul Rahman

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AB - 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. Conclusion: 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.

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