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
樊庭宇,曲戎梅,冯雁婷,彭彦,张仔昂,欧阳钧,戴景兴,钟世镇
中国临床解剖学杂志 ›› 2018, Vol. 36 ›› Issue (1) : 45-50.
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
目的 观察不同浓度细胞松弛素D(Cyto D)对人脂肪源干细胞(hASC)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、0.2 μg/ml的Cyto D处理hASC,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0.2 μg/ml的存活率相比于对照组下降了1/3。Cyto D能抑制hASCs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hASC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 1d, 4d and 7d. Results Cyto D can both induce the formation of lipid droplets and change the basal state of osteogenesis in hASC. 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.
脂肪源干细胞 / 成骨分化 / 成脂分化 / 肌动蛋白 / 细胞松弛素D
Adipose-derived stem cells;   / Osteogenesis;   / Adipogenesis;   / Actin;   / Cytochalasin D
[1] 陈平,曲戎梅,韩忠宇,等. 人脂肪源干细胞促进前交叉韧带重建后腱骨界面愈合的实验研究[J]. 中国临床解剖学杂志, 2016, 34(6):643-646.
[2] Tholpady SS, Llull R, Ogle RC, et al. Adipose tissue: stem cells and beyond[J]. Clinics in Plastic Surgery, 2006, 33(1):55-62.
[3] Zuk PA, Zhu M, Mizuno H, et al. Multilineage cells from human adipose tissue: implications for cell-based therapies[J]. Tissue Eng, 2001,7(2):211-228.
[4] 杜文佳,汪玉良,党跃修,等. 细胞松弛素D对大鼠脊髓星形胶质细胞水通道蛋白和内向整流性钾通道4.1基因表达的影响[J]. 中国康复理论与实践, 2014, 20(7):616-620.
[5] 陈波,唐康来,张吉强,等. 抑制肌动蛋白聚合对体外大鼠跟腱来源肌腱干细胞成脂分化的影响研究[J]. 中国修复重建外科杂志, 2015, 29(2):206-212.
[6] Sen B, Xie Z, Uzer G, et al. Intranuclear actin regulates osteogenesis[J]. Stem Cells, 2015, 33(10):3065-3076.
[7] Skarp K, Huet G, Vartiainen MK. Steady-state nuclear actin levels are determined by export competent actin pool[J]. Cytoskeleton, 2013, 70(10):623-634.
[8] 张扬,刘大诚,杨效宁. 不同培养条件下脂肪干细胞与成骨细胞的共培养[J]. 中国组织工程研究, 2014, 18(37):6003-6007.
[9] 陈犹白,陈聪慧,Zhang Qixu,等. 脂肪干细胞成骨分化的研究进展[J]. 中华损伤与修复杂志(电子版), 2016, 11(2):126-134.
[10] 陈犹白,郝永红,王岚,等. 脂肪干细胞成脂分化的分子机制和信号通路[J]. 中国组织工程研究, 2017, 21(1):154-158.
国家重点研发计划资助(2017YFC1105000)
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