钙离子浓度对脂肪干细胞增殖和成肌分化的影响  

Effects of Calcium Concentration on Proliferation and Myogenic Differentiation of Adipose Stem Cells

作　　者：陈皓[1] 柯晨 张梓凯 李力群[2] 林明 CHEN Hao;KE Chen;ZHANG Zi-kai;LILi-qun;LIN Ming(The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University,Wenzhou,Zhejiang,325000,China;The First Affiliated Hospital of Wenzhou Medical University,Wenzhou,Zhejiang,325000,China)

机构地区：[1]温州医科大学附属第二医院育英儿童医院,浙江温州325000 [2]温州医科大学附属第一医院,浙江温州325000

出　　处：《现代生物医学进展》2022年第7期1206-1210,共5页Progress in Modern Biomedicine

基　　金：温州市科技局市级基础性科研项目(Y20190017)。

摘　　要：目的:本研究旨在探讨钙离子浓度对脂肪干细胞(ADSCs)增殖和成肌分化的影响。方法:利用抽脂手术废弃脂肪提取脂肪干细胞,进行体外细胞培养和鉴定。设立正常细胞培养液为对照组,在培养液中添加不同浓度的钙离子(分别为0.001 mol/mL、0.002 mol/mL、0.003 mol/mL、0.004 mol/mL、0.005 mol/mL共5组)为实验组,测定在48 h和96 h条件下不同浓度钙离子(Ca^(2+))对cck8反应细胞增殖的影响。同时通过TGF-β诱导成肌分化,测定不同钙离子浓度下Ca^(2+)信号相关蛋白的表达,并通过CCK-8检测评价其分化水平。另外通过免疫荧光和PCR检测细胞表面特异性抗原(α-SMA,SMMHC)的表达水平。各组间比较应用独立样本t检验,P<0.05为差异具有统计学意义。结果:显微镜下观察可见原代ADSCs细胞培养0.5天后贴壁生长,形态呈圆或椭圆形,而TGF-β诱导ADSCs成肌分化可见细胞呈梭形;实验组中,钙浓度<0.002 mmol/mL时,脂肪干细胞增殖得到显著促进(t=10.08,df=10,P<0.05);钙浓度<0.003 mmol/mL时,脂肪干细胞的成肌分化也显著增加,同时免疫荧光和PCR检测发现低浓度下α-SMA,SMMHC的表达均显著增加(P<0.05),而高钙浓度下两者均受到抑制(P<0.05)。结论:低钙浓度(钙浓度<0.002 mmol/mL)可显著促进脂肪干细胞增殖和成肌分化,而高钙浓度下两者均受到抑制。Objective:The aim of this study was to investigate using myogenic differentiation of adipose stem cells for the treatment of female pelvic floor dysfunction(PFD)and aimed to investigate the effect of calcium concentration on the proliferation and myogenic differentiation of adipose stem cells.Methods:Human adipose stem cells were obtained from adipose tissue samples collected during plastic surgery and cultured.These were divided into six groups:one group was assigned as the blank control group,and the other five groups were added with different concentrations of calcium ions,respectively.Under the conditions of 48 and 96 hours,the proliferation of CCK8-reactive cells in different concentrations of Calcium ion(Ca^(2+))was measured.In the second part,under the induction of TGF-βmyogenic differentiation,different concentrations of Ca^(2+)were added to evaluate the myogenic differentiation process of proteins involved in Ca^(2+)signal under experimental conditions and control groups,and these were measured by CCK-8,immunofluorescence and PCR to determine the expression level of the cell surface specific antigen(α-SMA,SMMHC).Results:After0.5 days of culture,microscopic observation showed that ADSCs cells adhered to the wall and grew in round or oval shape.After the inducement of TGF-β,myogenic differentiation of ADSCs showed fusiform cells.In the experimental group,the proliferation of adipocytes was significantly promoted when calcium concentration was less than 0.002 mmol/m L(T=10.08,DF=10,P<0.05).When calcium concentration was less than 0.003 mmol/m L,the myogenic differentiation of adipocytes was also significantly increased,and the expression ofα-SMA and SMMHC were significantly increased at low calcium concentration(P<0.05),while they were inhibited at high calcium concentration(P<0.05).Conclusion:Myogenic differentiation and proliferation of adipose stem cells were also significantly increased in low calcium concentration(calcium concentration<0.003 mmol/mL),P<0.05,while both of them were inhibited in high

关 键 词：钙离子 脂肪干细胞 成肌分化 细胞增殖 表面特异性抗原 

分 类 号：R-33[医药卫生] Q26[生物学—细胞生物学]