The Effect of Betaine on Osteogenic Differentiation of Human Adipose-Derived Mesenchymal Stem Cells In Vitro

Objective: Human adipose- derived stem cells (hASCs) are af-fected by many biochemical and biophysical stimuli in vivo. Betaine (a dietary supplement) is a biochemical stimulant that promotes the osteogenic differentiation by increasing the entry of calcium into the cell. In this study, the effect of Betaine on the expression of osteogenic genes was evaluated.Material and Methods: hASCs were extracted from abdomi-nal adipose tissue, and stem cells were confirmed by flow cytometry. The experimental groups were: cultured cells in osteogenesis differentiation medium (positive control), in os-teogenesis differentiation medium and Betaine ۱۰ mM (BET + OD), in Betaine (BET) and negative control. On day ۲۱, alka-line phosphatase (ALP) activity and expression of osteogenic genes (RUNX۲ and OCN) were assessed by Real time PCR.Results: A significant increase in ALP activity in BET group was detected compared to OD + BET and positive control. On day ۷, there was a significant increase in ALP activity com-pared to ۱۴-day treatment. There was a significant decrease in the mRNA level of RUNX۲ in BET group compared to positive control. The expression of OCN gene in BET group was signifi-cantly increased compared to the OD + BET, while a significant increase of this gene was seen in positive control compared to OD + BET. There was no significant difference between BET and positive control.Conclusion: The induction of hADSCs with Betaine increased ALP activity and OCN gene expression and decreased RUNX۲ gene expression. The effect of Betaine on OCN gene expression was similar to osteogenesis differentiation medium. Betaine had effective role on ALP activity at an earlier time post induction.

Keywords:

Human Adipose-Derived Stem Cells , Betaine , Osteo-genic Genes , Real Time PCR ,