Stemness and angiogenic gene expression changes of serial-passage human amnion mesenchymal cells
| Autor: | A R Hayati, Ay Eeng Tan, Kien Hui Chua, Simat Siti Fatimah, Geok Chin Tan, Mohd Manzor Nur Fariha |
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| Rok vydání: | 2013 |
| Předmět: |
Adult
Angiogenesis Primary Cell Culture CD34 Neovascularization Physiologic Osteocytes Biochemistry Immunophenotyping Mesoderm Young Adult Pregnancy Adipocytes Humans CD90 Amnion Angiogenic Proteins Cells Cultured Neurons Neural Plate biology CD44 Mesenchymal stem cell Gene Expression Regulation Developmental Cell Differentiation Epithelial Cells Mesenchymal Stem Cells Cell Biology Fibroblasts Nestin Antigens Differentiation Molecular biology Culture Media Cell biology biology.protein Female Stem cell Cardiology and Cardiovascular Medicine Cell Division Fetal bovine serum |
| Zdroj: | Microvascular Research. 86:21-29 |
| ISSN: | 0026-2862 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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