title: Impact of adipogenic differentiation on stemness and osteogenic gene expression in extensive culture of human adipose-derived stem cells.
abstract: BACKGROUND
  Adipose tissue is a source of multipotent adult stem cells. Most studies on human adipose-derived stem cells (ASC) have been on the early passages. Studies in extensive expansion have not been well established yet. In this study, we aim to investigate the effects of extensive expansion on the adipogenic differentiation capability of ASC.
  
  
  METHODS
  The ability of ASC to undergo adipogenic differentiation in extensive expansion was evaluated by morphological changes, differentiation assay by using Oil Red O staining and changes in the genes expression levels of adipogenic genes, osteogenic genes and stemness genes using quantitative polymerase chain reaction (qPCR) after induction.
  
  
  RESULTS
  Morphological study showed that the formation of lipid droplets can be observed at all passages but decreased at P20 after induction. Data from qPCR showed that most adipogenicgenes expression increased significantlyat P5, P10 and P15 but decreased at P20 after induction. On the other hand, osteogenic genes showed no significant changes after adipogenic induction indicating low potentiality of adipogenic-induced ASC to become osteogenic cells. While stemness genes expression levels showed a decrease or no significant changes after adipogenic induction except Nanog3, which showed a significant increase at P15 and P20.
  
  
  CONCLUSIONS
  The ability of ASC to differentiate into mature adipogenic cells decreased after P10 and the decrease in the osteogenics gene expression level during adipogenic induction suggested that the osteogenesis and adipogenesis are not parallel events.
authors.affiliation: Department of Physiology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia.
authors.forename: Wan Kamarul Zaman Wan
authors.initials: WK
authors.lastname: Safwani
authors.affiliation: Department of Biochemistry, Faculty of Medicine, Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia.
authors.forename: Suzana
authors.initials: S
authors.lastname: Makpol
authors.affiliation: Subang Jaya Medical Centre, Selangor, Malaysia.
authors.forename: Somasundaram
authors.initials: S
authors.lastname: Sathapan
authors.affiliation: Department of Physiology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia.
authors.forename: Kienhui
authors.initials: K
authors.lastname: Chua
keywords: adipogenesis; adipose; cell-based therapy; culture expansion; differentiation; osteogenesis; stem cells; stemness
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