中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2012年
44期
3143-3146
,共4页
胡安斌%商昌珍%闵军%蔡继业%何晓顺
鬍安斌%商昌珍%閔軍%蔡繼業%何曉順
호안빈%상창진%민군%채계업%하효순
胚胎干细胞%上皮型钙黏蛋白%细胞分化%肝细胞
胚胎榦細胞%上皮型鈣黏蛋白%細胞分化%肝細胞
배태간세포%상피형개점단백%세포분화%간세포
Embryonic stem cells%Epithelial cadherin%Cell differentiation%Hepatocytes
目的 观察上皮型钙黏蛋白(E-cad)对小鼠胚胎干细胞(ESC)在体外向肝细胞分化的增强调控作用.方法 将E-cad转染入BALB/c小鼠ESC使其稳定表达,再利用肝细胞生长因子(HGF)等诱导其向肝细胞分化.然后用逆转录-聚合酶链反应(RT-PCR)检测肝细胞标记基因ALB、TAT、Cyp7a1的表达,用RIA法检测培养液ALB和尿素合成情况,显微镜观察分化细胞数量和黏附状态.结果 (1)普通ESC在拟胚体(EB)形成后表达E-cad最强,其后表达降低至17 d不再表达,而E-cad-ESC则稳定表达E-cad; (2)肝细胞标记基因ALB、TAT和Cyp7a1在E-cad-ESC组表达要早于或强于普通ESC组,ALB和尿素合成方面,E-cad-ESC组早于普通ESC组[普通ESC组,ALB最早出现于第11天,浓度为(0.20 ±0.03) mg/L,在17 d达到最高为(3.71±0.45)mg/L;尿素最早出现于第12天,浓度为(8.01±1.21) μmol/L,在17 d达到最高为(45.01±8.01)μmol/L.E-cad-ESC组,ALB最早出现于第9天,浓度为(0.19±0.03)mg/L,在17d达到最高为(5.52±0.82) mg/L ;尿素最早出现于第10天,浓度为(5.90±1.21) μmol/L,在17 d达到最高为(56.93±9.03)μmol/L],差异有统计学意义 ;(3)普通ESC在分化后期表现为松散的细胞群落,分化细胞数量稀少,E-cad-ESC分化细胞数量明显增多,细胞黏附能力增强并呈紧密的多层生长.结论 E-cad可增强ESC向肝细胞的分化作用,表现为分化细胞数量增多并呈多层生长,ALB及尿素合成能力增强,有望成为一种在ESC向肝脏方向分化研究中的重要调控因子.
目的 觀察上皮型鈣黏蛋白(E-cad)對小鼠胚胎榦細胞(ESC)在體外嚮肝細胞分化的增彊調控作用.方法 將E-cad轉染入BALB/c小鼠ESC使其穩定錶達,再利用肝細胞生長因子(HGF)等誘導其嚮肝細胞分化.然後用逆轉錄-聚閤酶鏈反應(RT-PCR)檢測肝細胞標記基因ALB、TAT、Cyp7a1的錶達,用RIA法檢測培養液ALB和尿素閤成情況,顯微鏡觀察分化細胞數量和黏附狀態.結果 (1)普通ESC在擬胚體(EB)形成後錶達E-cad最彊,其後錶達降低至17 d不再錶達,而E-cad-ESC則穩定錶達E-cad; (2)肝細胞標記基因ALB、TAT和Cyp7a1在E-cad-ESC組錶達要早于或彊于普通ESC組,ALB和尿素閤成方麵,E-cad-ESC組早于普通ESC組[普通ESC組,ALB最早齣現于第11天,濃度為(0.20 ±0.03) mg/L,在17 d達到最高為(3.71±0.45)mg/L;尿素最早齣現于第12天,濃度為(8.01±1.21) μmol/L,在17 d達到最高為(45.01±8.01)μmol/L.E-cad-ESC組,ALB最早齣現于第9天,濃度為(0.19±0.03)mg/L,在17d達到最高為(5.52±0.82) mg/L ;尿素最早齣現于第10天,濃度為(5.90±1.21) μmol/L,在17 d達到最高為(56.93±9.03)μmol/L],差異有統計學意義 ;(3)普通ESC在分化後期錶現為鬆散的細胞群落,分化細胞數量稀少,E-cad-ESC分化細胞數量明顯增多,細胞黏附能力增彊併呈緊密的多層生長.結論 E-cad可增彊ESC嚮肝細胞的分化作用,錶現為分化細胞數量增多併呈多層生長,ALB及尿素閤成能力增彊,有望成為一種在ESC嚮肝髒方嚮分化研究中的重要調控因子.
목적 관찰상피형개점단백(E-cad)대소서배태간세포(ESC)재체외향간세포분화적증강조공작용.방법 장E-cad전염입BALB/c소서ESC사기은정표체,재이용간세포생장인자(HGF)등유도기향간세포분화.연후용역전록-취합매련반응(RT-PCR)검측간세포표기기인ALB、TAT、Cyp7a1적표체,용RIA법검측배양액ALB화뇨소합성정황,현미경관찰분화세포수량화점부상태.결과 (1)보통ESC재의배체(EB)형성후표체E-cad최강,기후표체강저지17 d불재표체,이E-cad-ESC칙은정표체E-cad; (2)간세포표기기인ALB、TAT화Cyp7a1재E-cad-ESC조표체요조우혹강우보통ESC조,ALB화뇨소합성방면,E-cad-ESC조조우보통ESC조[보통ESC조,ALB최조출현우제11천,농도위(0.20 ±0.03) mg/L,재17 d체도최고위(3.71±0.45)mg/L;뇨소최조출현우제12천,농도위(8.01±1.21) μmol/L,재17 d체도최고위(45.01±8.01)μmol/L.E-cad-ESC조,ALB최조출현우제9천,농도위(0.19±0.03)mg/L,재17d체도최고위(5.52±0.82) mg/L ;뇨소최조출현우제10천,농도위(5.90±1.21) μmol/L,재17 d체도최고위(56.93±9.03)μmol/L],차이유통계학의의 ;(3)보통ESC재분화후기표현위송산적세포군락,분화세포수량희소,E-cad-ESC분화세포수량명현증다,세포점부능력증강병정긴밀적다층생장.결론 E-cad가증강ESC향간세포적분화작용,표현위분화세포수량증다병정다층생장,ALB급뇨소합성능력증강,유망성위일충재ESC향간장방향분화연구중적중요조공인자.
Objective To investigate the roles of E-cadherin (E-cad) in enhancing the in vitro differentiation of hepatocytes from murine embryonic stem cells (ESCs).Methods Exogenous E-cad was transfected into BALB/c murine ESCs to enable its stable expression.Then hepatic differentiation from E-cad-ESCs was induced by such growth factors as hepatocyte growth factor (HGF),fibroblast growth factor (FGF) and transforming growth factor (TGF).And the expressions of hepatic markers ALB,TAT,Cyp7a1and urea were detected.The morphology of hepatic differentiation was observed under microscopy.Results E-cad expression gradually decreased in normal ESC differentiation,but was stably expressed in E-cad-ESCs.In E-cad-ESC group,hepatic markers ALB,TAT and CYP7a1 were expressed earlier or higher than that in normal ESC group,and the concentrations of ALB and urea were significantly higher than that in normal ESC group.The adhesion of the differentiated E-cad-ESCs was significantly enhanced compared with the normal ESCs.They maintained close connections and multidimensional growth.Cell number of hepatocytes from ESC increased significantly in E-cad-ESC group.Conclusion E-cad enhances the hepatic differentiation of ESC by increasing the number of differentiated cells and increasing the synthetic capacity of ALB and urea.
