Figure 1: Characteristics of human adipose-derived stem cells (ASCs). Cultured ASCs were prepared from stromal vascular fraction cells isolated from collagenase-digested human subcutaneous adipose tissue. The cells were analyzed for their multilineage differentiation potential in vitro. (A) Morphology of ASCs at first passage. (B) Cells were cultured in adipogenic differentiation medium for 2 weeks and stained with Oil red O. (C and D) Cells were cultured in osteogenic differentiation medium for 3 weeks and stained for alkaline phosphatase (ALP) (C) and alizarin red S (D). (E and F) Cells were seeded in 15-cm3 conical tubes and cultured in chondrogenic differentiation medium for 3 weeks. Formed micromass pellets were photographed (E). The micromass pellets were sectioned and stained for aggrecan (F). (G) Cells were cultured in smooth muscle cell differentiation medium for 1 week and stained for α-smooth muscle actin (ASMA). Nuclei were stained with Hoechst 33342 (Hoechst). Scale bar: 200 μm in A, B, C, D, F, and G; 1 mm in E.