A comparative study on two culture methods for adipose-derived stem cells from mice with osteoporosis / 中国组织工程研究

ABSTRACT

BACKGROUND:To obtain enough adipose-derived stem cells (ADSCs) is the premise of repairing osteoporotic bone defects by autograft transplantation;therefore,how to efficiently,simply and economically harvest primary autologous ADSCs is the key to the treatment of osteoporosis.OBJECTIVE:To isolate and culture ADSCs from mice with osteoporosis (OP-ADSCs) in vitro by tissue explant culture and collagenase digestion,and to compare the efficacies of these two methods.METHODS:Adipose tissues were isolated from the inguen of C57BL/6 mice,from which OP-ADSCs were obtained by tissue explant culture and collagenase digestion respectively.Then,the cells were subjected to primary culture and subculture.The surface specific antigens of passage 3 cells were observed using flow cytometry,while the yields and proliferation abilities of passage 3 were compared at 7 days of culture.The adipogenic and osteogenic differentiation of OP-ADSCs at passage 4 was detected.RESULTS AND CONCLUSION:(1) The expression levels of CD34,CD146,Sca-1 in passage 3 cells were (15.22±1.85)％,(75.55±3.36)％ and (83.48±4.22)％ for the tissue explant culture and (13.46±2.21)％,(76.62±2.47)％ and (84.84±3.56)％ for the collagenase digestion method,respectively.(2) The cell yield from each milligram of adipose tissue by tissue explant culture was significantly higher than that by collagenase digestion (P ＜ 0.05).(3) The proliferation rate of cells in the tissue explant culture group was higher than that in the collagenase digestion group after 24 hours of culture.(4) The OP-ADSCs cultured by the two methods could differentiate into adipocytes and osteoblasts,and the lipid accumulation and the mineralized nodules showed no significant difference between the two groups (both P＞ 0.05).These experimental findings indicate that the tissue explant culture is more suitable for obtaining OP-ADSCs in vitro as compared with collagenase digestion,which contributes to provide adequate cell sources for studies on ADSCs treatment of osteoporotic fractures and bone defects.