Effects of Serial Passaging on the Adipogenic and Osteogenic Differentiation Potential of Adipose-Derived Human Mesenchymal Stem Cells

Adipose-derived human mesenchymal stem cells (hMSCs) will be more valuable for tissue engineering applications if they can be extensively subcultured without loss of phenotype and multilineage differentiation ability. This study examined the effects of serial passaging on growth rate, gene expressio...

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Bibliographic Details
Published in:Tissue engineering 2007-06, Vol.13 (6), p.1291-1298
Main Authors: Wall, Michelle E., Bernacki, Susan H., Loboa, Elizabeth G.
Format: Article
Language:English
Subjects:
Adipocytes - cytology
Adipocytes - physiology
Adipogenesis - physiology
Adult
Cell culture
Cell Culture Techniques - methods
Cell Differentiation
Cells, Cultured
Cellular biology
Female
Gene expression
Humans
Mesenchymal Stromal Cells - cytology
Mesenchymal Stromal Cells - physiology
Middle Aged
Osteocytes - cytology
Osteocytes - physiology
Osteogenesis - physiology
Stem cells
Tissue engineering
Tissue Engineering - methods
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Summary:Adipose-derived human mesenchymal stem cells (hMSCs) will be more valuable for tissue engineering applications if they can be extensively subcultured without loss of phenotype and multilineage differentiation ability. This study examined the effects of serial passaging on growth rate, gene expression, and differentiation potential of adipose-derived hMSCs. Differentiation was assessed by analyzing changes in messenger RNA (mRNA) expression of osteogenic and adipogenic marker genes and by determining production of calcium deposits and lipid vacuoles. Cells cultured in osteogenic medium for 2 weeks upregulated expression of alkaline phosphatase mRNA relative to cells in growth medium, and deposited calcium. Calcium deposition decreased in cells from passages 4 to 6 but returned to levels near or above those of primary cells by passage 10. Cells cultured in adipogenic medium upregulated expression of lipoprotein lipase and peroxisome proliferator activated receptor-γ mRNA relative to cells in growth medium, and formed lipid vacuoles at all passages. By passage 8, however, cells in adipogenic medium also deposited calcium. Growth rate was stable through passage 5, then decreased. The results of this study indicate that adipose-derived hMSCs are capable of both adipogenic and osteogenic differentiation through 10 passages (34 population doublings) but that osteogenic differentiation may start to dominate at later passages.
ISSN:1076-3279
1557-8690
DOI:10.1089/ten.2006.0275