不同浓度细胞松弛素D对人脂肪源干细胞分化能力的影响  

The effect of different concentrations of cytochalasin D on the differentiation ability of human adipose-derived stem cells

作　　者：樊庭宇 曲戎梅[1] 冯雁婷 彭彦 张仔昂 欧阳钧[1] 戴景兴[1] 钟世镇[1] FAN Ting-yu;QU Rong-mei;FENG Yan-ting;PENG Yan;ZHANG Zi-ang;OUYANG Jun;DAI Jing-xing;ZHONG Shi-zhen(Guangdong Provincial Key Laboratory of Construction and Inspection in Tissue Engineering, Department of Anatomy, Southern Medical University, Guangzhou 510515, China)

机构地区：[1]南方医科大学人体解剖教研室暨广东省组织构建与检测重点实验

出　　处：《中国临床解剖学杂志》2018年第1期45-50,共6页Chinese Journal of Clinical Anatomy

基　　金：国家重点研发计划资助(2017YFC1105000)

摘　　要：目的观察不同浓度细胞松弛素D（Cyto D）对人脂肪源干细胞（h ASC）成骨分化和成脂分化能力的影响。方法在生长培养基、成脂诱导培养基及成骨诱导培养基中分别加入0.05、0.1、0.2μg/ml的Cyto D处理h ASC,干扰肌动蛋白的延长,并在处理的第1、4、7 d时分别进行油红O染色、ALP染色及CCK8等实验检测细胞性能的改变。结果 7 d时实验组0.2μg/ml的存活率相比于对照组下降了1/3。Cyto D能抑制h ASCs的存活和增殖,且浓度越大细胞的存活率越低。单纯的Cyto D既能促使h ASC胞内脂滴形成,又能改变细胞内成骨的基础状态。Cyto D与成脂诱导液联合使用后能极大的促进成脂,且不同的浓度具有不同的成脂效果,低浓度使脂滴数量增加,高浓度促进脂滴成熟;与成骨诱导液联合使用后Cyto D用药浓度和成骨效果成反比。结论细胞松弛素D能增强成骨或成脂诱导液效果,且低浓度细胞松弛素D能够促进成骨,低浓度和高浓度细胞松弛素D都能促进成脂。本实验探究了细胞松弛素D引起的肌动蛋白解聚对脂肪源干细胞的分化能力影响,为研究脂肪源干细胞的分化机制提供了重要生物学信息。Objective To observe the effect of different concentrations of cytochalasin D（Cyto D）on the differentiation ability of human adipogenic stem cells to osteogenesis and adipogenesis. Methods Different concentrations of Cyto D that 0.05 μg/ml, 0.1μg/ml and 0.2μg/ml were added to growth medium,osteogenic differentiation medium and adipogenic differentiation medium, respectively, to disturb the elongation of cytoskeletal actin in different environments. Then the changes of cell differentiation ability were detected by oil red O staining, ALP staining and CCK8, in the treatment of 1 d, 4 d and 7 d. Results Cyto D can both induce the formation of lipid droplets and change the basal state of osteogenesis in h ASC. The combination of Cyto D and adipogenic differentiation medium can greatly promote the lipids formation. Low concentrations Cyto D increased the number of lipid droplets, and high concentrations Cyto D promoted the maturation of fat droplets. In combination with osteogenic differentiation medium, Cyto D was inversely proportional to the concentration and osteogenic effect. Conclusion Cyto D can enhance the effect of differentiation medium. Low concentration Cyto D mainly promotes osteogenesis and high concentration of Cyto D mainly promotes adipogenesis. Our exploration on the effect of myosin on the differentiation of adipogenic stem cells induced by Cyto D can provide important biological information for studying the differentiation mechanism of adipogenic stem cells.

关 键 词：脂肪源干细胞 成骨分化 成脂分化 肌动蛋白 细胞松弛素D 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]