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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 a decentralized culture model of human decidual cells for screening anti-fertility drugs. Methods Normal human decidua tissues from 6 to 9 weeks of early pregnancy were digested with collagenase (150u / ml) and hyaluronidase (50u / ml) in a constant temperature water bath at 35 ℃ for 100min and filtered through 216μm stainless mesh Of human decidua stromal cells; the cells were cultured in serum-free medium (hereinafter referred to as the culture medium), by changing the culture medium to add growth factors and anti-fertility drug concentrations observed growth changes. Results Human decidual cells in serum-free FD + 8F culture can maintain adherent growth more than 3 weeks. Epidermal growth factor, insulin, transferrin, selenium, hydrocortisone and small doses of progesterone (10-8mol / L) and estradiol (10-8mol / L) have the effect of maintaining the growth of decidual cells; Reproductive material yuanhuacine (0.5μg / ml), mifepristone (Ru486,10μg / ml), tamoxifen (10μg / ml), hyaluronic acid (50μg / ml) Ester (60μg / ml) had a killing effect on cultured human decidual cells, while TC had no significant effect (100μg / ml). Mifepristone combined with tamoxifen and anordrin, respectively, have a synergistic effect on the injury of decidual cells. Conclusion Human decidual cell culture has many advantages as a primary screening model of anti-fertility drugs and needs further development and promotion.