Marshburn P B, Head J R, MacDonald P C, Casey M L
Cecil H. and Ida Green Center for Reproductive Biology Sciences, Obstetrics and Gynecology, University of Texas Southwestern Medical Center, Dallas 75235-9032.
Am J Obstet Gynecol. 1992 Dec;167(6):1888-98. doi: 10.1016/0002-9378(92)91792-9.
The purpose of this study was to evaluate the culture characteristics and cell proliferation of human endometrial glandular epithelial cells in primary culture on extracellular matrix and to evaluate sex steroid modulation of this process.
We examined the culture characteristics in 53 endometrial gland preparations obtained throughout the menstrual cycle and determined the incorporation of tritiated thymidine in endometrial epithelial cells as a measure of deoxyribonucleic acid synthesis (cell proliferation) in the presence and absence of 17 beta-estradiol and medroxyprogesterone acetate.
Good culture maintenance of endometrial epithelial monolayers on extracellular matrix was observed from glands derived from proliferate phase endometrium (21 of 23), whereas poor adherence and culture maintenance was observed in all 30 of specimens from the secretory phase. With 17 beta-estradiol treatment for 22 hours, tritiated thymidine incorporation was not different from control, but medroxyprogesterone acetate treatment for 22 hours was associated with diminished tritiated thymidine incorporation by 36% (p < 0.004). When 17 beta-estradiol (10(-8) mol/L) was included in the incubation medium from the time of initial culture, tritiated thymidine incorporation on day 4 was 40% of control (p < 0.006), and tritiated thymidine was not suppressed further by 22 hours of treatment with medroxyprogesterone acetate (10(-7) mol/L). In spite of cellular spread of endometrial epithelial cells to subconfluence, deoxyribonucleic acid content per well did not increase over time in culture.
We conclude that exposure of endometrial glands to progesterone in vivo inhibits adherence and the establishment of cell monolayers cultured on a thin layer of extracellular matrix. Because 17 beta-estradiol treatment in culture for 22 hours does not increase tritiated thymidine incorporation, it is possible that 17 beta-estradiol exerts its proliferative effect on the endometrial epithelium by an indirect action mediated by stromal cells. The effects of 17 beta-estradiol and medroxyprogesterone acetate on deoxyribonucleic acid synthesis in endometrial epithelial cells may represent an action on a stem cell population of dividing cells or a terminally differentiated cell population that undergoes programmed cell death.
本研究旨在评估人子宫内膜腺上皮细胞在细胞外基质上原代培养时的培养特性和细胞增殖情况，并评估性激素对这一过程的调节作用。
我们检查了在整个月经周期获取的53份子宫内膜腺体标本的培养特性，并在有和没有17β-雌二醇及醋酸甲羟孕酮存在的情况下，测定氚标记胸腺嘧啶核苷掺入子宫内膜上皮细胞的情况，以此作为脱氧核糖核酸合成（细胞增殖）的指标。
观察到来自增殖期子宫内膜的腺体在细胞外基质上能良好维持子宫内膜上皮单层培养（23份中有21份），而来自分泌期的所有30份标本的贴壁和培养维持情况较差。用17β-雌二醇处理22小时后，氚标记胸腺嘧啶核苷掺入情况与对照无差异，但用醋酸甲羟孕酮处理22小时后，氚标记胸腺嘧啶核苷掺入减少了36%（p<0.004）。当从初始培养时就在孵育培养基中加入17β-雌二醇（10⁻⁸mol/L）时，第4天的氚标记胸腺嘧啶核苷掺入量为对照的40%（p<0.006），且用醋酸甲羟孕酮（10⁻⁷mol/L）处理22小时后，氚标记胸腺嘧啶核苷未进一步受到抑制。尽管子宫内膜上皮细胞铺展至亚汇合状态，但培养中每孔的脱氧核糖核酸含量并未随时间增加。
我们得出结论，子宫内膜腺体在体内暴露于孕酮会抑制在薄层细胞外基质上培养的细胞单层的贴壁和形成。由于在培养中用17β-雌二醇处理22小时不会增加氚标记胸腺嘧啶核苷掺入，因此17β-雌二醇可能通过基质细胞介导的间接作用对子宫内膜上皮发挥增殖作用。17β-雌二醇和醋酸甲羟孕酮对子宫内膜上皮细胞脱氧核糖核酸合成的影响可能代表对正在分裂的干细胞群体或经历程序性细胞死亡的终末分化细胞群体的作用。
