摘要
目的:比较人羊膜上皮细胞(human amnion epithelial cells,h AECs)与人羊膜间充质干细胞(human amnion mesenchymal stem cells,h AM SCs)的生物学特性。方法:从足月剖宫产胎盘上剥离羊膜组织,采用低速胰蛋白酶-胶原酶消化法,分别分离获取h AECs及h AMSCs,观察此两种细胞的增殖生长情况,采用免疫荧光染色、流式细胞术、逆转录聚合酶链反应(reverse transcription-polymerase chain reaction,RT-PCR)、诱导分化染色等方法对两种细胞进行生物学特性鉴定及对比。结果:h AECs为圆形或椭圆形,呈铺路石样生长,最多传至5代;h AMSCs为成纤维细胞样的长梭形,呈放射状或漩涡状分布生长,可传至30代左右。h AECs表达上皮细胞标志角蛋白19(cytokeratin19,CK19),部分表达波形蛋白;h AM SCs表达波形蛋白,不表达CK19。两种细胞均表达CD29、CD44、CD73、CD90、OCT-4、Nanog。h AECs不表达CD105,成骨及成脂诱导阳性。结论:h AECs和h AM SCs均具有某些干细胞特性,h AECs不能无限增殖,其生长增殖及分化能力均低于h AM SCs。
Objective : To compare the biological characteristics between the human amnion epithelial cells( h AECs)and human amnion mesenchymal stem cells( h AM SCs). Me thods: Human amnion tissues were peeled off from human full term placenta from the women with normal healthy singleton pregnancies undergoing elective cesarean section. Using lowspeed-trypsin collagenase digestion method,h AECs and h AM SCs were isolated from human amnion tissue. The proliferation and biological characteristics of both cells were analyzed and compared by immunofluorescence assay,flowcytometry,reverse transcription-polymerase chain reaction( RT-PCR),induced differentiation and so on. Re sults:h AECs were round or oval,and formed a confluent monolayer of cobblestone-shaped epithelial cells. h AECs continue to proliferate less than 5 passages,while the h AM SCs were fibroblastic and spindle-shaped,which could continue to proliferate about 30 passages. h AECs expressed CK19 and a fewof them expressed vimentin; h AM SCs expressed vimentin and did not express CK19. Both h AECs and h AM SCs were positive for suface markers CD29,CD73,CD44,CD90,OCT-4,Nanog and so on,but h AECs were negative for CD105. Both h AECs and h AM SCs were positive for oil red O and akane red staining. Conclusions: h AECs and h AM SCs display stem cell properties. h AECs can not proliferate unlimitly,and the proliferation and differentiation ability of h AECs is lower than h AM SCs.
出处
《中国实验血液学杂志》
CAS
CSCD
北大核心
2015年第4期1120-1124,共5页
Journal of Experimental Hematology
关键词
人羊膜上皮细胞
人羊膜间充质干细胞
生物学特性
human amnion epithelial cells
human amnion mesenchymal stem cells
biological characteristic
