In vitro proliferation and differentiation of adipose-derived stem cells isolated using anti-CD105 magnetic beads

Abstract

The present study aimed to investigate the feasibility of isolating adipose-derived stem cells (ADSCs) by selecting cells that express the surface receptor CD105. Surface antigen expression of the unsorted cells was undertaken using FACS analysis. Primary adipose-derived cells were isolated. The second passage cells were incubated with anti-CD105 magnetic beads, and separated using a magnetic separator. Cell growth and colony formation was determined by counting and Giemsa staining, respectively. Cells also underwent histological immunohistochemical, and RT-PCR analyses to determine their chondrogenic, adipogenic and osteogenic potential. Increased cell proliferation and colony formation was observed in CD105-positive (CD105+) as compared to the CD105-negative (CD105-) cells (P<0.001). Following induction, the expression of type II collagen and the number of calcium deposits and lipid droplets in the CD105+ ADCs were markedly higher than in the CD105- ADCs. Furthermore, increased alkaline phos phatase (AKP), leptin and PPARγ2 mRNA expression was detected in the CD105+ ADCs (P<0.01). Isolation of CD105+ ADSCs by MACS was feasible. Thus, CD105 can be used as a relatively specific marker for the selection of ADSCs. Although the chondrogenic, adipogenic and osteogenic potential of these cells is suggestive of their potential for use in tissue engineering treatments, further in vivo studies are necessary.