摘要
目的探讨脂肪源性干细胞(ADSCs)及其分化的施万细胞(SCs)在形态、活性和标志物表达上的区别。方法取SD大鼠腹股沟处脂肪制备ADSCs,用流式细胞术检测细胞表面标志;钙钴金属盐沉淀法检测碱性磷酸酶(ALP),茜素红染色观察矿化结节;油红O染色观察脂滴;随机分对照组和诱导组,加入条件培养基向SCs诱导分化;MTT法检测诱导后第2、4、6和8 d各时间点的细胞活性;扫描电镜及免疫染色观察S100的表达。结果光镜下可见ADSCs呈梭形贴壁生长;ADSCs表达抗原分化簇90(CD90)和抗原分化簇105(CD105),不表达抗原分化簇45(CD45);成骨诱导后可见ALP呈强阳性表达,茜素红染色可见橘红色矿化结节,油红O染色可见胞内红染脂滴聚集;向SCs诱导后第4 d,可见多数细胞呈纺锤形,第8 d纺锤状细胞增多;MTT检测显示第4 d诱导组的吸光度明显低于对照组,差异有统计学意义(P<0.05);组内相比,诱导组和对照组的吸光度均随着时间的延长而逐渐增大,差异有统计学意义(P<0.05);第6、第8 d诱导组与对照组吸光度差异无统计学意义(P>0.05);扫描电镜可见诱导后第8 d细胞细长,细胞外成分多,S100阳性表达;ADSCs表面呈山脊状,纤毛密集,细胞外成分少,不表达S100。结论ADSCs与其分化的SCs形态差异显著,分化而来的SCs活性良好,表达SCs标志蛋白。
Objective To explore the differences in morphology,activity and expression of markers of adipose-derived stem cells(ADSCs)and their differentiated Schwann cells(SCs).Methods Flow cytometry detection of cell surface markers;Determination of alkaline phosphatase(ALP)by calcium and cobalt metal salt precipitation method,alizarin red staining to observe mineralized nodules;Observation of lipid droplets by oil red O staining;Add conditioned medium to induce differentiation into SCs;MTT detection of cell viability at various time points on the2nd,4th,6th,and 8th days after induction;Scanning electron microscope observation and immunocytochemical staining detection expression S100.Results Observation under light microscope shows that ADSCs were fusiform;ADSCs expressed CD90 and CD105,but not CD45;Strong positive expression of ALP can be seen after osteoinduction,alizarin red staining shows orange-red mineralized nodules,Oil Red O staining shows intracellular lipid droplet aggregation;On the 4th day after induction to SCs,most of the cells were spindle-shaped,increased spindle cells on the8th day;MTT test showed that the absorbance of the induction group was significantly lower than that of the control group on the 4th day,and the difference was statistically significant(P<0.05);Compared within the group,the absorbance of the induction group and the control group gradually increased with time,and the difference was statistically significant(P<0.05);There was no statistically significant difference in absorbance between the induction group and the control group on the 6th and 8th day(P>0.05);Scanning electron microscopy showed that on the 8th day after induction,the cells were slender and had many extracellular components,and S100 protein was positively expressed.The surface of ADSCs is ridged,with dense cilia,few extracellular components,and no S100 protein expression.Conclusion The morphological difference between ADSCs and their differentiated SCs is significant,the differentiated SCs have good activity and can express
作者
郑玲
邓佳茜
胡莹
刘杏
管海辰
饶利兵
鞠晓军
ZHENC Ling;DENG Jiaxi;HU Ying;LIU Xing;GUAN Hai-chen;RAO Li-bing;JU Xiao-jun(Clinical Skills Training Center,Hunan University of Medicine,Huaihua,Hunan Province 418000,China;Department of Human Anatomy,Hunan University of Medicine,Huaihua,Hunan Province 418000,China;Department of Imaging Diagnosis,Hunan University of Medicine,Huaihua,Hunan Province 418000,China)
出处
《解剖学研究》
CAS
2023年第5期405-410,共6页
Anatomy Research
基金
湖南省教育厅科学研究项目(湘教通[2022]323号-22B1033)
湖南省教育厅大学生创新创业训练计划项目(湘教通[2020]191号-4272)
湖南省中医药科研计划项目(湘中医药[2021]2号-2021095)。
关键词
脂肪源性干细胞
诱导分化
施万细胞
大鼠
Adipose-derived stem cells
Induced differentiation
Schwann cells
Rat
