Yeger H, Lines L D, Wong P Y, Silver M M
Department of Pathology, Hospital for Sick Children, Toronto, Ontario, Canada.
Placenta. 1989 Mar-Apr;10(2):137-51. doi: 10.1016/0143-4004(89)90036-2.
A simple method is described for the isolation of trophoblast cells from both first trimester and term placenta. Trophoblast preparations were characterized by light microscopy, scanning and transmission election miscroscopy and immunohistochemistry to distinguish these cells from mesenchyme and endothelium. Trophoblast cells were cultured on various substrates and a comparison made of their ability to attach, proliferate and function. A collagen gel substrate produced by repolymerization of an acid soluble collagen fraction from chorionic villi allowed rapid attachment of trophoblast cells and maintainance of their original morphology. Term trophoblast cells were shown to become fully functional in short term (three day) cultures by virtue of their increased immunocytochemical staining for the presence of beta hCG, hPL and SPI. beta hCG increased significantly by day three thus demonstrating functional activation. Trophoblast cells from first trimester placenta formed proliferating colonies of hormone producing cells while those from term placenta reaggregated into clusters and closely resembled syncytiotrophoblast both morphologically and functionally. This short term culture system for term trophoblast will allow further studies into the biology of trophoblast polypeptide hormone synthesis and secretion.
本文描述了一种从孕早期和足月胎盘分离滋养层细胞的简单方法。通过光学显微镜、扫描和透射电子显微镜以及免疫组织化学对滋养层细胞制剂进行表征，以将这些细胞与间充质和内皮细胞区分开来。将滋养层细胞培养在各种基质上，并比较它们附着、增殖和发挥功能的能力。由绒毛膜绒毛中酸溶性胶原部分重新聚合产生的胶原凝胶基质能够使滋养层细胞快速附着并保持其原始形态。足月滋养层细胞在短期（三天）培养中显示出通过β-hCG、人胎盘催乳素（hPL）和SPI的免疫细胞化学染色增加而变得完全有功能。到第三天β-hCG显著增加，从而证明了功能激活。孕早期胎盘的滋养层细胞形成了产生激素的细胞增殖集落，而足月胎盘的滋养层细胞重新聚集形成簇，在形态和功能上都与合体滋养层非常相似。这种用于足月滋养层细胞的短期培养系统将有助于对滋养层多肽激素合成和分泌的生物学进行进一步研究。
