人子宫内膜细胞体外诱导蜕膜化的效果观察

Comparison of Three Methods on Decidualization of Human Endometrial Cells in Vitro

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(P<0.05)。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。

Objective： To observe the effect of three different methods on decidualization of human endometrial cells in vitro. Methods： Human endometrial tissues were obtained from women undergoing hysterectomies for uterine benign diseases. Endometrium stromal cells （ESCs） were separated and cul- tured in vitro and identified by immunofluorescence. The 3rd generation of ESCs were treated with progestogen （P） ＋ estrogen （E2）（PE group）, P＋8-Br-cAMP （PC group） and P＋E2＋8-Br-cAMP （PEC group）, respectively. The cell morphology was observed under the microscope, and the PRL level of cell-culture medium in 24 h, 48 h and 96 h were detected using chemiluminescence method. Results： ESCs became round with bigger cell volume and nucleus, and the cell boundaries became ambiguous during decidualization. Among these three different groups, cells treated with P＋8-Br-cAMP showed more obvious changes in both PRL level and cell morphology. Conclusion： Compared with P＋E2 and P＋E2＋8-Br-cAMP, P＋8-Br-cAMP is a more effective method on induction of decidualization of endometrial cells in vitro.