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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 effects of different combinations of progestogen (P), estrogen (E_2) and 8-Br-cAMP on human endometrial cells induced decidual The effect of the Methods: Endometriosis was collected from patients with benign uterine disease. The endometrial stromal cells (ESCs) were isolated and purified. The cells were cultured in vitro and subcultured. The cell types were identified by immunofluorescence. The third generation of ESC was induced by three different combinations of P + E_2 (PE group), 8-Br-cAMP (PC group) and P + E_2 + 8-BrcAMP (PEC group), respectively. For the control group. The morphological changes of cells during the process of decidualization were observed under a microscope. The levels of prolactin (PRL) in the culture medium were detected by chemiluminescence assay after 24h, 48h and 96h respectively by three different decidualization induction methods. Three decidualization methods were compared Effect. Results: During the process of decidualization, the morphology of ESC gradually changed from long fusiform to round shape. The size of the cells increased, the nuclei increased, some nuclei or nuclei appeared, and the cell borders became blurred. With the prolongation of culture time, the levels of PRL in each decidua induction group increased at different levels. The levels of PRL and decidualized cells in PC group were significantly higher than those in PE and PEC groups (P <0.05). Conclusion: The P + 8-Br-cAMP method is more effective than the P + E_2 and P + E_2 + 8-BrcAMP methods in inducing the decidualization of the uterine ESC in a more efficient manner, which is an efficient and reliable method to induce decidualization.