人羊膜上皮细胞向卵泡样结构转分化的实验研究

发布时间：2018-01-12 01:17

本文关键词：人羊膜上皮细胞向卵泡样结构转分化的实验研究　出处：《四川大学学报(医学版)》2017年04期 　论文类型：期刊论文

【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(P0.05),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(P0.05)。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。
[Abstract]:Objective to study human amniotic epithelial cells in human amniotic epithelial cells. HAECs could be transformed into follicle-like structure after long time stimulation of human follicular fluid. Methods on the basis of the preliminary results, 5% human follicular fluid was isolated and cultured in vitro. The induction time was extended to 44 days. The morphological changes of cells were observed by inverted microscope and the level of estradiolium E2 in the supernatant of culture medium was detected by chemiluminescence immunoassay. Real-time fluorescence quantification-polymerase chain reaction real time-polymerase chain reaction. The female germ cell specific gene (GDF9) was detected by RT-PCR. Results in the induction group (medium containing 5% follicular fluid), the oocyte like cells (OLCs) appeared on the 10th day of culture, and the volume of OLCs increased with time. Around the transparent zona-like ring change, then decreased, partially disappeared; The expression of mRNA in DAZL and GDF9 in the induced group was higher than that in the control group (P 0.05) and had two peaks. There was no significant difference in the expression of SCP3mRNA between the induction group and the control group (P 0.05). Conclusion hAECs have the potential to differentiate into follicle-like structure in vitro.
【作者单位】：四川大学华西第二医院妇产科;出生缺陷与相关妇儿疾病教育部重点实验室(四川大学);陕西省人民医院;
【基金】：四川省科技厅科技支撑计划项目(No.2011SZ0137)资助
【分类号】：R339.2
【正文快照】： 近年来,妇科恶性肿瘤在世界范围内的发病率越来越高,且出现年轻化趋势[1-2]。妇科恶性肿瘤常用的治疗仍以手术为主,再辅以放化疗,治疗后最为突出的并发症为卵巢结构或功能受损所致的不孕和低雌激素水平相关的围绝经期综合征。为解决这些问题,亟需寻求一种与体内雌激素效价、代

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