两种方法培养骨质疏松症小鼠脂肪干细胞的比较
A comparative study on two culture methods for adipose-derived stem cells from mice with osteoporosis
摘要
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.关键词
干细胞/脂肪干细胞/骨质疏松症/组织块培养法/胶原酶消化法/成脂分化/成骨分化/国家自然科学基金分类
医药卫生引用本文复制引用
黄成龙,黎庆,王雷,黄馗,罗道文,肖金刚..两种方法培养骨质疏松症小鼠脂肪干细胞的比较[J].中国组织工程研究,2017,21(29):4654-4659,6.基金项目
国家自然科学基金项目(81371125) （81371125）
四川省科技厅项目(2014JY0044) （2014JY0044）
四川省教育厅项目(10ZB030) （10ZB030）
四川省卫生厅项目(80170) （80170）
西南医科大学重点项目(201207) （201207）
2015年泸州市人民政府-西南医科大学科技战略合作项目[2015LZCYD-S05(2/12)] （2/12）
西南医大口院[(2017)20号] （2017）
the National Natural Science Foundation of China,No.81371125 （）
the Project of Sichuan Science and Technology Department,No.2014JY0044 （）
the Project of Sichuan Education Department,No.10ZB030 （）
the Project of Sichuan Health Department,No.80170 （）
the Major Project of Southwest Medical University,No.201207 （）
the Science and Technology Strategic Cooperation Project of Luzhou Municipal People's Government-Southwest Medical University,No.2015LZCYD-S05(2/12) （2/12）
a grant from the Hospital of Stomatology of Southwest Medical University,No.(2017)20 （2017）
