Figure 1.

Characterization of a novel human ASCL. (A) Schema shows the process to establish an ASCL from human subcutaneous adipose tissue. Adipose tissue (0.5-1 g) digested with collagenase was centrifuged to obtain ASCs. ASCs were cultured in conditioned media to differentiate into mature adipocytes. Fat drops from trypsinized cells were moved to an upside-down culture. Floating mature adipocytes with fat droplets were collected and placed into the flask, which was completely filled with the media. In the flask completely filled with medium over 14 days without medium change, mature adipocytes with a fat droplet on the flask surface changed into the ASCL. A morphological change from a fat drop into adherent cells indicates that an ASCL is obtained. (B) A proliferation assay was performed on an ASCL established from 3 subjects. (C) The ASCL differentiated into osteoblasts, mature adipocytes, and chondrocytes. The ASCL was cultured in conditioned media for each differentiation, and we assessed their capacity for differentiation using cell staining. (D) Expression of surface markers in the ASCL was estimated by flow cytometry. (E) Representative data for karyotype analysis of the ASCL.