Tiemann U, Hansen P J
Forschungsinstitut für die Biologie landwirtschaftlicher Nutztiere, Dummerstorf, Germany.
In Vitro Cell Dev Biol Anim. 1995 Sep;31(8):640-5. doi: 10.1007/BF02634318.
Cultured cells from the bovine endosalpinx were used to evaluate effects of estradiol-17 beta, progesterone, epidermal growth factor, and insulinlike growth factors I and II on [3H]thymidine incorporation. Cells were treated with hormones and growth factors when approximately 50% confluent. After 24 h, DNA synthesis was quantified by pulsing cells with [3H]thymidine for 12 h and determining uptake into DNA. Cells prepared by mechanical dispersal incorporated more [3H]thymidine than cells dispersed with collagenase. However, hormonal responses were the same for both types of cells. As compared to plastic, cells on a Matrigel substratum exhibited lower incorporation of [3H]thymidine and were unresponsive to hormones. Estradiol-17 beta increased [3H]thymidine incorporation slightly at 10(-10) mol/liter and higher. Epidermal growth factor, insulinlike growth factor-I, and insulinlike growth factor-II also stimulated [3H]thymidine incorporation. Effects of insulinlike growth factor-I were greater for cells treated with estradiol-17 beta. In the absence of estradiol, progesterone inhibited [3H]thymidine incorporation at 1, 10, and 100 ng/ml. When estradiol-17 beta was present, progesterone stimulated [3H]thymidine incorporation at 1 ng/ml and reduced incorporation at 100 ng/ml. In conclusion, [3H]thymidine incorporation by cultured oviductal endosalpingeal cells can be regulated by ovarian steroids and growth factors. These molecules may represent signals through which the ovary, embryo, and oviduct regulate oviductal growth.
利用牛输卵管内膜的培养细胞来评估17β-雌二醇、孕酮、表皮生长因子以及胰岛素样生长因子I和II对[3H]胸腺嘧啶核苷掺入的影响。当细胞达到约50%汇合时,用激素和生长因子进行处理。24小时后,通过用[3H]胸腺嘧啶核苷脉冲细胞12小时并测定其掺入DNA的量来定量DNA合成。机械分散制备的细胞比用胶原酶分散的细胞掺入更多的[3H]胸腺嘧啶核苷。然而,两种类型的细胞对激素的反应是相同的。与塑料相比,基质胶上的细胞[3H]胸腺嘧啶核苷掺入率较低,且对激素无反应。17β-雌二醇在10^(-10)摩尔/升及更高浓度时略微增加[3H]胸腺嘧啶核苷掺入。表皮生长因子、胰岛素样生长因子-I和胰岛素样生长因子-II也刺激[3H]胸腺嘧啶核苷掺入。胰岛素样生长因子-I对用17β-雌二醇处理的细胞的作用更大。在没有雌二醇的情况下,孕酮在1、10和100纳克/毫升时抑制[3H]胸腺嘧啶核苷掺入。当存在17β-雌二醇时,孕酮在1纳克/毫升时刺激[3H]胸腺嘧啶核苷掺入,而在100纳克/毫升时降低掺入。总之,培养的输卵管内膜细胞的[3H]胸腺嘧啶核苷掺入可受卵巢甾体激素和生长因子调节。这些分子可能代表卵巢、胚胎和输卵管调节输卵管生长的信号。