中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2015年
6期
1323-1328
,共6页
人羊水克隆样细胞来源干细胞%Transwell%3-D联合共培养%肝样细胞
人羊水剋隆樣細胞來源榦細胞%Transwell%3-D聯閤共培養%肝樣細胞
인양수극륭양세포래원간세포%Transwell%3-D연합공배양%간양세포
Human amniotic fluid colony-derived stem cells%Transwell%3-D co-culture%Hepatocyte-like cells
目的 探讨Transwell 3-D联合共培养促使人羊水克隆样细胞来源干细胞向肝样细胞分化的可能性.方法 利用克隆杯,从人孕中期羊水标本中分离获得人羊水克隆样细胞来源干细胞,培养扩增后,通过流式细胞仪检测表面抗原进行鉴定.利用Transwell小室联合共培养肝细胞诱导其向肝样细胞分化,观察诱导后细胞形态的变化,利用细胞免疫荧光检测甲胎蛋白(AFP)、白蛋白(ALB)和细胞角蛋白-18(CK-18)在诱导后细胞中的表达,反转录-聚合酶链反应(RT-PCR)检测肝细胞相关基因的表达.结果 利用克隆杯分离出的人羊水克隆样细胞来源干细胞CD44和CD90表达阳性,CD10、CD14、CD34、CD45和人类白细胞抗原DR基因(HLA-DR)表达阴性.诱导2～4周后,肝样细胞出现,检测证实这些细胞表达AFP、ALB、CK-18、GATA结合蛋白-4(GATA4)、肝细胞核因子(HNF)-1α和细胞色素20A(CYP20A)等肝细胞相关标志和基因.结论 Transwell 3-D联合共培养可以促使人羊水克隆样细胞来源干细胞向肝样细胞分化.
目的 探討Transwell 3-D聯閤共培養促使人羊水剋隆樣細胞來源榦細胞嚮肝樣細胞分化的可能性.方法 利用剋隆杯,從人孕中期羊水標本中分離穫得人羊水剋隆樣細胞來源榦細胞,培養擴增後,通過流式細胞儀檢測錶麵抗原進行鑒定.利用Transwell小室聯閤共培養肝細胞誘導其嚮肝樣細胞分化,觀察誘導後細胞形態的變化,利用細胞免疫熒光檢測甲胎蛋白(AFP)、白蛋白(ALB)和細胞角蛋白-18(CK-18)在誘導後細胞中的錶達,反轉錄-聚閤酶鏈反應(RT-PCR)檢測肝細胞相關基因的錶達.結果 利用剋隆杯分離齣的人羊水剋隆樣細胞來源榦細胞CD44和CD90錶達暘性,CD10、CD14、CD34、CD45和人類白細胞抗原DR基因(HLA-DR)錶達陰性.誘導2～4週後,肝樣細胞齣現,檢測證實這些細胞錶達AFP、ALB、CK-18、GATA結閤蛋白-4(GATA4)、肝細胞覈因子(HNF)-1α和細胞色素20A(CYP20A)等肝細胞相關標誌和基因.結論 Transwell 3-D聯閤共培養可以促使人羊水剋隆樣細胞來源榦細胞嚮肝樣細胞分化.
목적 탐토Transwell 3-D연합공배양촉사인양수극륭양세포래원간세포향간양세포분화적가능성.방법 이용극륭배,종인잉중기양수표본중분리획득인양수극륭양세포래원간세포,배양확증후,통과류식세포의검측표면항원진행감정.이용Transwell소실연합공배양간세포유도기향간양세포분화,관찰유도후세포형태적변화,이용세포면역형광검측갑태단백(AFP)、백단백(ALB)화세포각단백-18(CK-18)재유도후세포중적표체,반전록-취합매련반응(RT-PCR)검측간세포상관기인적표체.결과 이용극륭배분리출적인양수극륭양세포래원간세포CD44화CD90표체양성,CD10、CD14、CD34、CD45화인류백세포항원DR기인(HLA-DR)표체음성.유도2～4주후,간양세포출현,검측증실저사세포표체AFP、ALB、CK-18、GATA결합단백-4(GATA4)、간세포핵인자(HNF)-1α화세포색소20A(CYP20A)등간세포상관표지화기인.결론 Transwell 3-D연합공배양가이촉사인양수극륭양세포래원간세포향간양세포분화.
Objective To investigate the potential of human amniotic fluid colony-derived stem cells (hAFCSCs) differentiated into hepatocyte-like cells using transwell co-culture.Methods Using colony poculum,hAFCSCs were isolated from second-trimester amniotic fluid.After cultured and amplified,the expression of surface antigens was identified by flow cytometry.The hAFCSCs were induced into hepatocyte-like cells using transwell co-culture.Observe the change of cell morphology after co-culture.The expression of alpha-fetoprotein (AFP),Albumin (ALB),Cytokeratin-18 (CK-18) and hepatocyte gene was detected with immunofluorescence and reverse transcriptase-polymerase chain reaction (RT-PCR).Results hAFCSCs were positive for CD44,CD90,and negative for CD10,CD14,CD34,CD45 and human leukocyte antigen-DR (HLA-DR).After 2-4 weeks induction,hepatocyte-like cells appeared.It confirmed that these cells expressed AFP,ALB,CK-18,GATA4,Hepatocyte nuclear factor (HNF)-1 α and cytochrome 20A (CYP20A).Conclusion Human amniotic fluid colony-derived stem cells can be differentiated into hepatocyte-like cells using transwell co-culture.
