人子宫内膜细胞体外诱导蜕膜化的效果观察

发布时间：2018-02-28 16:39

本文关键词： 子宫内膜细胞蜕膜化 -溴-环磷酸腺苷(-Br-cAMP) 雌激素(E_) 孕激素(P)　出处：《生殖与避孕》2014年01期 　论文类型：期刊论文

【摘要】：目的:观察孕激素(progestogen,P)、雌激素(estrogen,E_2)和8-溴-环磷酸腺苷(8-Br-cAMP)不同组合的3种方法对人子宫内膜细胞体外诱导蜕膜化的效果。方法:收集因子宫良性疾病行全子宫切除术患者的子宫内膜,分离纯化子宫内膜间质细胞(endometrial stromal cell,ESC),进行细胞体外培养并传代,免疫荧光鉴定细胞类型。分别采用P+E_2(PE组)、8-Br-cAMP(PC组)、P+E_2+8-BrcAMP(PEC组)3种不同组合方法蜕膜化诱导处理第3代ESC,并以空白对照为对照组。显微镜下观察蜕膜化过程中的细胞形态变化,化学发光法检测3种不同蜕膜化诱导方法处理24h.48h及96h后细胞培养液中催乳素(PRL)水平,比较3种蜕膜化方法的效果。结果:蜕膜化过程中ESC形态从长梭形逐渐变为圆形,细胞体积变大,胞核增大,部分出现双核或多核,细胞边界变模糊。随着培养时间延长,各蜕膜化诱导组培养液中PRL呈不同水平升高。其中PC组PRL水平及蜕膜化细胞形态均较PE组和PEC组升高明显(P0.05)。结论:P+8-Br-cAMP法,较P+E_2、P+E_2+8-BrcAMP法能更有效地诱导子宫ESC体外发生蜕膜化,是高效可靠、更合适的蜕膜化诱导方法。
[Abstract]:Objective: to observe the effects of three different combinations of progestogen P (P), estradiol estradiol (E _ 2) and 8-Br-cAMP) on decidualization induced by human endometrial cells in vitro. Methods: total hysterectomy was performed for benign uterine diseases. The endometrium of the patient, Endometrial stromal cells were isolated and purified from endometrial stromal cells, cultured in vitro and subcultured. The cell types were identified by immunofluorescence. The third generation ESCs were treated by decidualization induction with three different combinations of P E2PE group and 8-Br-cAMPP PC group. The morphologic changes of the cells during decidualization were observed under microscope. Chemiluminescence assay was used to detect the prolactin prolactin (PRL) levels in the cell culture medium after 24 h, 48 h and 96 h treatment with different decidualization methods, and to compare the effects of the three decidualization methods. Results: during decidualization, the shape of ESC gradually changed from spindle shape to round shape. The size of the cell becomes larger, the nucleus of the cell increases, part of the cell has binuclear or polynuclear, and the boundary of the cell becomes blurred. The PRL level and decidualized cell morphology in PC group were significantly higher than those in PE group and PEC group (P 0.05). Conclusion the ratio of P 8-Br-cAMP method is more effective than P E2P E28 Br-BrcAMP method in inducing decidualization of uterine ESC in vitro. It is an effective and more suitable method for decidualization induction.
【作者单位】：首都医科大学附属北京妇产医院生殖中心;首都医科大学附属北京妇产医院妇科;首都医科大学附属北京妇产医院手术室;首都医科大学附属北京妇产医院检验科;
【基金】：北京市自然科学基金项目(5122015) 北京市科技新星基金项目(H020821200190) 首都医科大学附属北京妇产医院院内基金项目(201114)
【分类号】：R329

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