人羊膜上皮细胞向卵泡样结构转分化的实验研究  

Trans-differentiation of Human Amniotic Epithelial Cells into Follicle-like Structure

作　　者：王慧[1,2] 王飞[3] 白莉平[1,2] 莫小雨 唐慧芸 郑艾[1,2] 

机构地区：[1]四川大学华西第二医院妇产科,成都610041 [2]出生缺陷与相关妇儿疾病教育部重点实验室(四川大学),成都610041 [3]陕西省人民医院,西安710000

出　　处：《四川大学学报（医学版）》2017年第4期531-536,共6页Journal of Sichuan University(Medical Sciences)

基　　金：四川省科技厅科技支撑计划项目(No.2011SZ0137)资助

摘　　要：目的研究人羊膜上皮细胞(human amniotic epithelial cells,hAECs)在人卵泡液的长时间刺激下能否转分化为卵泡样结构。方法在前期实验结果基础上,体外分离培养hAECs,5%人卵泡液为诱导条件,延长诱导时间至44d,倒置显微镜观察细胞形态学改变,化学发光免疫技术检测培养基上清液中雌激素(estradiol,E2)水平的变化,实时荧光定量-聚合酶链式反应(real time-polymerase chain reaction,RT-PCR)检测雌性生殖细胞特异性基因(GDF9、DAZL、SCP3)表达情况。结果诱导组(添加有5%卵泡液的培养基)在培养第10天出现卵母细胞样细胞(OLCs),随着时间延长,OLCs体积增大,周围见透明带样环变,随后减小,部分消失;对照组无上述现象。诱导组DAZL及GDF9的mRNA表达水平较对照组升高(P<0.05),并且具有两个高峰;诱导组SCP3mRNA表达水平与对照组之间的差异无统计学意义(P>0.05)。诱导组有E2生成,其水平随时间先下降后升高再下降;对照组无E2生成。结论在体外,hAECs具有向卵泡样结构转分化的潜能。Objective To determine whether hAECs can differentiate into follicle-like structure. Methods hAECs were isolated from human term placenta and cultured sequentially in medium supplemented with 5 % human follicular fluid. The morphology of the cells was examined by inverted microscope. Changes of estradiol levels and expressions of germ cell specific gene in the cultured medium were detected by chemiluminescence immunoassay and Real time-Polymerase Chain Reaction （RT-PCR）, respectively. Results Oocyte-like cells （OLCs） were found in those cultured in the medium supplemented with 5% human follicular fluid. The OLCs gradually aggrandized, shrunk and finally disappeared over time. However, this phenomenon was not observed in the control group. Higher expressions of DAZL and GDF9 were found in the follicle group compared with the control which medium without 5%human follicular fluid＇ （P〈0.05）, with two peaks. No significant difference in the expression of SCP3 was found between the two groups （P〈0.05）. Estradiol was detected in the culture medium of the follicle group in a mode of decreasing-rising-decreasing, but not in the control group. Conclusion hAECs have the potential to trans-differentiate into the follicle-like structure in vitro.

关 键 词：人羊膜上皮细胞 人卵泡液 转分化 雌性生殖细胞 卵泡样结构 雌激素 

分 类 号：R339.2[医药卫生—人体生理学]