[Culture of the single cell clones of human adipose-derived stem cells and identification of their surface antigen expressions].

OBJECTIVE:To perform single cell clone culture of human adipose-derived stem cells (hASCs), and identify their surface antigens in different clones. METHODS:Surgically removed human adipose tissues were digested with collagenase to isolate hASCs, which were seeded in primary culture. After culture to the second passage, the hASCs were committed to limiting dilution assays to form the colony units. Each of the clones obtained were examined with flow cytometry to identify the expressions of cell-surface antigens including CD29, CD44, CD34, CD54, CD106, and ABCG2. RESULTS:Ten clones were obtained by colony-forming unit (CFU) assays, and after culture to the 9th passage, a portion of the cells were frozen in liquid nitrogen. All the clones exhibited similar fibroblast-like morphology, but their proliferative activities varied. Flow cytometry showed that all the clones were strongly positive for CD29 and CD44 but with rather low ABCG2 expression. The expressions of CD34, CD54 and CD106 varied between the cell clones. CONCLUSION:hASCs harvested from human adipose tissue with collagenase digestion represent a hybrid population of multilineage stem cells. CFU assays may result in single purified hASC clones, and the different clones differ in but also share some common surface antigens, possibly due to their different potentials of differentiation.