Abstract : Adipose-derived stem cells (ADSCs) are promising stem cell for autologous regenerative therapy. However, culturing process of ADSCs manipulated the size of cells into senescence formed. The aim of this study was to investigate the effect of concentration variation of ascorbic acid towards proliferation of human ADSCs. Human ADSCs were isolated by enzymatic digestion and cultured in presence serum. To evaluate trilineage ability of ADSCs, multipotency assay into adipocyte, chondrocyte and osteocyte were conducted. The prepared cells then cultured into 8 groups involved cells cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) only, DMEM with 10% foetal bovine serum (FBS) (DMFA), DMFA with 25, 50, 75, 100, 125, and 150 µg/ml of ascorbic acid, respectively. Morphology observation including spread size of cells, population doubling time, viability and proliferation curve were performed. Analysis of type 1 and type 2 collagen and vascular endothelial growth factor (VEGF) as well were conducted in mRNA level using quantitative-real time PCR. The results showed that the cells had plastic adherence capacity and could be differentiated into adipocyte, chondrocytes, and osteocyte. After 120 hours of observation, the cells which cultured with FBS without ascorbic acid supplementation has the highest spread cells in size, lowest in cell yield and lowest in doubling time while the lowest spread size cells with the highest of cell yield and fastest in doubling time was in group with 75 µ

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