Douglas G C, King B F
Department of Human Anatomy, School of Medicine, University of California, Davis 95616.
In Vitro Cell Dev Biol. 1990 Aug;26(8):754-8. doi: 10.1007/BF02623616.
Trophoblast is the major functional cell type of the placenta. The purpose of this study was to devise a means to isolate trophoblast cells from the monkey placenta and to examine their capacity to differentiate in vitro. Methods originally devised for the isolation of human cytotrophoblast cells produced poor yields and a low degree of purity when applied to the near-term rhesus monkey placenta. However, a procedure has been developed which allows the isolation of a cell population consisting of more than 95% cytotrophoblast based on intermediate filament immunocytochemistry. The cells sedimented between densities of 1.040 and 1.053 g/ml on continuous Percoll density gradient centrifugation. When maintained in culture the cells adhered and formed aggregates of mononuclear cells by 24 h. By 5 d in culture, immunofluorescent staining using antidesmoplakin and antinuclear antibodies revealed that most colonies consisted of large multinucleated masses similar to syncytiotrophoblast. These results demonstrate trophoblast cells from monkey placental villi can be isolated with a high degree of purity and undergo morphologic differentiation in vitro. This preparation should enable investigators to study many functional characteristics of these cells throughout gestation.
滋养层细胞是胎盘的主要功能细胞类型。本研究的目的是设计一种从猴胎盘中分离滋养层细胞的方法，并检测其体外分化能力。最初设计用于分离人细胞滋养层细胞的方法，应用于近足月恒河猴胎盘时，产量低且纯度不高。然而，现已开发出一种方法，基于中间丝免疫细胞化学可分离出细胞滋养层细胞占比超过95%的细胞群体。通过连续Percoll密度梯度离心，细胞沉淀在密度为1.040至1.053 g/ml之间。在培养过程中，细胞在24小时内贴壁并形成单核细胞聚集体。培养5天时，使用抗桥粒斑蛋白抗体和抗核抗体进行免疫荧光染色显示，大多数集落由类似于合体滋养层的大型多核团块组成。这些结果表明，猴胎盘绒毛的滋养层细胞可以高度纯化地分离出来，并在体外发生形态学分化。这种制备方法应能使研究人员在整个妊娠期研究这些细胞的许多功能特性。
