Abstract:Objective To observe the proliferation capacity and adipogenic ability of mouse adipose-derived stem cells (mASCs) on different passages in vitro and to provide experimental evidence for rational selection of seed cells. Methods The mouse inguinal fat tissues were digested with collagenase, and cells were adhered to dishes, passage-cultured. The mASCs were divided into three groups: primary cells (P0), the second generation cells (P2) and the fifth generation cells (P5), and cell morphology and proliferation capacity of different passages of mASCs were observed. After four weeks of non-induction culture with high density seeding, the spontaneous adipogenic ability was observed through oil red staining and PT-PCR analysis. After two weeks of adipogenic induction, the adipogenic potential of mASCs was observed through oil red staining and PT-PCR analysis. Results The mouse adipose-derived stem cells displayed fibroblast-like morphology or irregular shape and different passages of them all had strong proliferation capacity. Primary cells had extremely strong spontaneous adipogenic ability in vitro. The spontaneous adipogenic ability of the second generation cells obviously reduced, and there was hardly any spontaneous adipogenic ability at passage 5. P0, P2 and P5 had similar induced adipogenic potential. Conclusion With the generation increasing, spontaneous adipogenic ability of mASCs reduced, but induced adipogenic potential almost unchanged. The fifth generation cells still maintain strong proliferation capacity and properties of stem cells. In a certain range of passage and amplification could reduce the content of pre-adipocyte and help purify the mesenchymal stem cells.

Key words:Mouse,Adipose-derived stem cells,Different passages,Adipogenic differentiation