人蜕膜细胞培养作为抗早孕药物筛选模型的研究

Study of Human Decidual Cell Culture as A Model for Screening Drugs to Terminate Early Pregnancy

目的建立分散的人蜕膜质细胞培养模型以用于抗生育药物的筛选研究。方法早孕６～９周的正常人蜕膜组织经胶原酶（１５０ｕ／ｍｌ）和透明质酸酶（５０ｕ／ｍｌ））于３５℃恒温水浴中振荡消化１００ｍｉｎ，并经２１６μｍ不锈钢网过滤后得到分离的人蜕膜基质细胞；该细胞培养于无血清培养液（以下简称培液）中，通过变换培液中添加生长因子及抗生育药物的浓度观察其生长变化情形。结果人蜕膜细胞在无血清ＦＤ＋８Ｆ培液中，可维持贴壁生长３周以上。上皮生长因子、胰岛素、转铁蛋白、硒、氢化可的松及小剂量的孕酮（１０－８ｍｏｌ／Ｌ）和雌二醇（１０－８ｍｏｌ／Ｌ）有维持蜕膜细胞生长的作用；抗生育物质芫花酯甲（０．５μｇ／ｍｌ）、米非司酮（Ｒｕ４８６，１０μｇ／ｍｌ）、他莫西芬（１０μｇ／ｍｌ）、土槿皮乙酸（５０μｇ／ｍｌ）和双炔失碳酯（６０μｇ／ｍｌ）对于培养的人蜕膜细胞均有杀伤作用，而天花粉（１００μｇ／ｍｌ）无明显作用。米非司酮与他莫西芬、双炔失碳酯分别合并使用，对蜕膜细胞的损伤有协同作用。结论人蜕膜细胞培养作为抗生育药物初筛模型有较多优点，有待进一步发展推广。

Objective To establish an in vitro model of human decidual cell culture for screening drugs and studying antifertility drugs.Methods Normal decidual tissues at 6th 9th weeks of gestation were minced and digested for 100 min with coll. (150u/ml) and hyalu. (50u/ml) at 35℃ constantly. The resulting supernatant was filtered through the stainless steel sieve of 216 μm to obtain isolated decidual cells. The decidual cells were cultured in serum free medium. The growth and change of the cells were observed as concentrations of added growth factors or drugs were being changed.Results Human decidual cells which were cultured in serum free FD+8F medium could be kept alive for over 3 weeks. EGF, INS, TF, Se, Hydroc and low dose (10 -8 mol/L) of P and E 2 could maintain the growth of decidual cells. The results showed that Yuanhuacine (0.5μg/ml), Ru486(10μg/ml), Tamoxifen (10μg/ml), Pseudolaric acid B (50μg/ml) and Anordrin (60μg/ml) were detrimental to the decidual cells but Trichosanthin had no evident effects. Coordinative effects were observed when Ru486 was given together with Tamoxifen or Anordrin respectively.Conclusion Human decidual cell culture has more advantages as a model for screening antifertility drugs but it deserves further research to develop.