羊水来源胎儿间充质干细胞定向诱导为脂肪细胞的研究  

The induction of fetal mesenchymal stem cells derived from amniotic fluid into adipocytes

作　　者：潘骏[1] 李现铎[2] 朱哲[2] 耿红全[2] 谢华[2] 陈方[2] 

机构地区：[1]上海市儿科医学研究所 [2]上海交通大学医学院附属新华医院泌尿外科,上海200092

出　　处：《中国临床解剖学杂志》2007年第1期64-67,共4页Chinese Journal of Clinical Anatomy

基　　金：国家自然科学基金资助项目(30271351)

摘　　要：目的:研究体外诱导孕中期羊水来源胎儿间质干细胞(M SCs)分化为脂肪细胞。方法:机械手段挑取孕中期羊水细胞培养体系中的胎儿M SCs集落,培养扩增后,通过流式细胞仪检测表面抗原表达进行鉴定。取第3代胎儿M SCs,IBM X、胰岛素、氢化考的松等诱导其向脂肪细胞分化。倒置相差显微镜观察诱导后细胞形态的变化。诱导20 d后,油红O染色检测胞内中性脂肪并计算阳性细胞率,W estern Blot蛋白印记法检测PPA Rγ的表达。结果:机械手段分离出的胎儿M SCs为CD 44阳性,CD 34、CD 45、H LA-D R阴性,符合M SCs特点。诱导72 h后,细胞内有脂滴出现,随着时间延长,脂滴逐渐增加并融合为脂泡,细胞由梭形转变为圆形或多角性。20 d后85%以上胎儿M SCs变为油红O染色阳性,W estern Blot显示诱导20 d后,诱导组细胞PPA Rγ的表达量明显高于未诱导组。结论:经上述方法诱导20 d后,羊水来源胎儿M SCs可以分化为脂肪细胞。Objective： To investigate the possibility of human fetal mesenchymal stem cells （MSCs） derived from second-trimester amniotic fluid being induced into adipocytes. Methods： Fibroblast-like fetal MSCs were separated mechanically from cultured amniocytes, expanded and analyzed its surface antigens by flow cytometry. After 3 passages, fetal MSCs were shifted to adipogenic medium （DMEM supplemented with FBS, IBMX, insulin, hydrocortisone, etc） for 20 days. The morphology of induced cells were evaluated by contrast microscopy. The rate of positive cells including intracellular lipid droplets was confirmed by Oil Red O staining. Western Blot was used to study the expression of PPARγ. Results： Fetal MSCs could be isolated from the culture of amniocytes using mechanical methods. The expanded MSCs were positive for CD44, but negative for CD34, CD45 and HLA-DR, that was consistent with the regular characters of MSCs line. After induction by adipogenic medium for 72 h, lipid droplets could be found in kytoplasm of MSCs. Over 85% fetal MSCs presented typical morphological feature of adipocytes, proved by positive Oil Red O staining results after 20 days induction. Western Blot showed the expression of PPARγ in the induced group was remarkable higher than that in the control group. Conclusions： Fetal MSCs derived from amniotic fluid can be induced into adipocytes after 20 days special induction.

关 键 词：羊水细胞 间充质干细胞 脂肪细胞 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]