胎盘滋养细胞微绒毛膜和基底膜的制备

Sequential Preparation of Microvillous and Basal Membranes from Human Placenta

目的改良制备胎盘滋养细胞微绒毛膜(microvillous membranes,MVM)和基底膜(basal membranes,BM)的技术,建立一个研究母胎界面物质交换的细胞分子模型。方法在Illsley法基础上改良缓冲液成分,延长镁离子螯合BM的时间,低速离心后的上清液用于制备MVM,沉淀用蔗糖梯度纯化制备BM。结果每克胎盘制备的MVM平均量为0.55 mg,其标记酶碱性磷酸酶(alkaline phosphatase,AKP)为空白对照的16.87倍;BM的平均量为0.54 mg,其标记酶腺苷酸环化酶(adenylate cyclase,AC)为空白对照的11.19倍。结论改良的Illsley法可以简单地同时制备较高质量的MVM和BM,有利于在细胞分子水平更好地研究胎盘滋养细胞功能。

Objective To improve the technology of isolating paired fractions of the maternal-facing membranes（MVM） and fetal-facing plasma membranes（BM） from a term placenta.Methods The component of buffer was improved based on Illsley method.The time of Mg^2＋-aggregated basal membranes was extended.MVM were obtained from the supernatant of low speed centrifugation while BM were further purified on a sucrose step gradient.Results Yield for MVM and BM prepared by the method were（0.55±0.10） mg/g and（0.54±0.02） mg/g wet weight of placenta.They were enriched 16.87-fold and 11.19-fold as determined by the membrane marker enzymes,alkaline phosphatase（MVM） and adenylate cyclase（BM）.Conclusion The modified Illsley method can easily produce both MVM and BM of satisfied quantity from human placenta.It could be applied as a cell molecular model of maternal-fetal exchange interface.