诱导人羊膜上皮细胞横向分化为肝细胞样细胞(英文)  

Transversal inducing differentiation of human amniotic epithelial cells into hepatocyte-like cells

作　　者：罗宏武[1] 黄湘俊[1] 黄飞舟[1] 刘浔阳[1] 

机构地区：[1]中南大学湘雅三医院肝胆外科,长沙410013

出　　处：《中南大学学报（医学版）》2011年第6期525-531,共7页Journal of Central South University ：Medical Science

摘　　要：目的:通过体外诱导分化实验,评价人羊膜上皮细胞(hAECs)向肝细胞样细胞分化的能力。方法:采用地塞米松(Dex),HGF,IGF等细胞因子联合诱导hAECs向肝细胞样细胞分化,诱导周期为2周,诱导过程中采用RT-PCR鉴定albumin(ALB),CYP1A1,CYP1A2,IGFR,c-met等肝细胞相关关键功能基因及HNF3,HNF4和C/EBPa 3种转录因子的表达,流式细胞术分析集落细胞表面标记ALB,AFP和CK18的时程变化。结果:诱导2周后hAECs表达ALB,CYP1A1,CYP1A2,c-met等肝细胞功能基因及HNF3,HNF4,C/EBPa,HNF1等转录因子,且功能基因随诱导天数的增加其表达逐渐增强;细胞表面标记检测发现:诱导第6天时,hAECs主要表达AFP+,为(15.1±2.1)%;随着诱导时间的延长,第10天时hAECs表达AFP+/ALB+,为(6.5±1.4)%;而诱导第14天时,hAECs基本上只表达ALB+,为(13.9±2.3)%。诱导第10天时的hAECs开始表达ALB+/CK18+;为(2.5±1.4)%;诱导第14天时,细胞依然表达ALB+/CK18+,且这种双阳性的细胞量有明显增加,为(18.9±3.1)%;同时CK18+细胞数并没有明显减少,诱导第10天时,CK18+细胞数为(16.1±1.2)%;诱导第14天时,CK18+细胞数为(21.3±4.6)%。提示诱导过程中的hAECs随着诱导时间的延长有一个逐渐成熟的过程。结论:体外诱导的hAECs具有分化为肝细胞样细胞的特性。Objective To evaluate the in vitro differentiation of human amniotic epithelial cells（hAECs） into hepatocyte-like cells.Methods Combined approach of dexamethasone,HGF,IGF and other cytokines were used to induce the differentiation of hAECs into hepatocyte-like cells.The induction lasted 2 weeks.During the induction,the expression of albumin ALB,CYP1A1,CYP1A2,IGFR,c-met and key functional genes related to liver cells as well as transcription factors HNF3,HNF4 and C/EBPa were monitored by RT-PCR.Time dependent changes of the surface marker colony ALB,AFP and CK18 were analyzed by cell flow cytometry.Results After the 2 week induction,the expressions of liver hepatocyte-like cell functional genes such as albumin,CYP1A1,CYP1A2,c-met,and transcription factors such as HNF3,HNF4,C/EBPa and HNF1 were observed.Six days after the induction,hAECs mainly were stained AFP＋,and the positive rate was（15.1±2.1）%.While 10 days after the induction,part of the hAECs showed AFP＋/ALB＋（6.5±1.4）%;and on 14th day,hAECs only showed ALB＋,and the rate was（13.9±2.3）%.ALB＋ cell increase indicated a gradual functional maturation from the hAECs to hepatocyte-like cells.Similaritly,the number of CK18＋ cells in the whole population was also increased： On 10th day,the rate was（16.1±1.2）%;on 14th day,that was（21.3±4.6）%,which proved the above hypothesis of the trandifferentiation.By extending the induction time,the expression of functional genes increased gradually,and a maturing process of hAECs was detected by cell surface markers.Conclusion The differentiation of hAECs induced in vitro has the characteristics of hepatocyte-like cells.

关 键 词：人羊膜上皮细胞 肝细胞样细胞 诱导分化 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]