不同血清体积分数对脂肪干细胞成脂分化的影响  

Effect of serum concentrations on differentiation of adipose-derived stem cells into adipocytes

作　　者：杨朵 李娜 Yang Duo;Li Na(Central Laboratory,Beijing Shijitan Hospital of Capital Medical University,Beijing 100038,China)

机构地区：[1]首都医科大学附属北京世纪坛医院中心实验室,北京市100038

出　　处：《中国组织工程研究》2018年第17期2669-2673,共5页Chinese Journal of Tissue Engineering Research

摘　　要：背景:培养基中不同体积分数血清对干细胞增殖分化有不同影响。目的:探讨含不同体积分数胎牛血清的培养基对小鼠脂肪干细胞增殖及成脂分化能力的影响。方法:取7-10 d龄C57小鼠皮下脂肪提取脂肪干细胞。培养基中胎牛血清体积分数设定为5%,10%,15%,20%,将小鼠脂肪干细胞培养至第3代,采用CCK-8法测定细胞的增殖能力。成脂诱导分化后采用Bodipy染色观察脂滴形成情况,采用荧光定量PCR法和western blot法检测Cebpa,Pparg基因表达量和C/EBP-α,PPAR-γ蛋白表达量。结果与结论:(1)体积分数为20%胎牛血清组脂肪干细胞在2-6 d的450 nm处吸光度值较体积分数为5%,10%,15%胎牛血清组低,但各组间差异无显著性意义;(2)体积分数为10%胎牛血清组Cebpa,Pparg基因表达量和C/EBP-α,PPAR-γ蛋白表达量最高,体积分数为5%,10%胎牛血清组显著高于体积分数为15%,20%胎牛血清组;(3)结果提示,不同体积分数胎牛血清培养条件对小鼠脂肪干细胞的增殖影响无显著差异。但不同体积分数胎牛血清培养条件下,脂肪干细胞成脂相关基因表达不同,高体积分数胎牛血清可抑制脂肪干细胞向脂肪细胞的分化。BACKGROUND： Serum concentrations in culture medium influence the proliferation and differentiation of stem cells. OBJECTIVE： To study the proliferation and adipogenic differentiation potential of adipose-derived stem cells under culture with different concentrations of fetal bovine serum.METHODS： The mouse adipose-derived stem cells were harvested from subcutaneous adipose tissue of C57 mice at 7-10 days of age. Concentrations of fetal bovine serum in the culture medium were set to 5%, 10%, 15% and 20%. The passage 3 cells were used for subsequent studies. The proliferation of cells cultured with different serum concentrations was tested using the cell counting kit-8 assay. The adipogenic differentiation potential of cells was evaluated with Bodipy staining. Quantitative RT-PCR and western blot assay were used to determine expression levels of Cebpa and Pparg m RNA as well as C/EBP-α and PPAR-γ protein levels. RESULTS AND CONCLUSION： There was no statistical difference in the cell proliferation among cells cultured with different serum concentrations, even though adipose-derived stem cells cultured with 20% fetal bovine serum showed lower absorbance value at 450 nm than those cultured with 5%, 10% and 15% fetal bovine serum. The expression levels of Cebpa and Pparg m RNA and C/EBP-α and PPAR-γ protein in the 5% and 10% groups were considerably higher than those of the 15% and 20% groups, and the expression levels were highest in the 10% group. To conclude, different concentrations of fetal bovine serum have no significant effects on the proliferation of adipose-derived stem cells, while the adipogenic differentiation potentials of cells are different when cultured with different serum concentrations. High serum concentration can inhibit the adipogenic differentiation of adipose-derived stem cells.

关 键 词：脂肪组织 干细胞 成脂分化 细胞培养技术 组织工程 脂肪干细胞 小鼠 血清 培养基 增殖 

分 类 号：R394.2[医药卫生—医学遗传学]