Faria T N, Soares M J
Department of Physiology, University of Kansas Medical Center, Kansas City 66103.
Endocrinology. 1991 Dec;129(6):2895-906. doi: 10.1210/endo-129-6-2895.
The purpose of this investigation was to establish and characterize a cell line derived from a rat choriocarcinoma and to evaluate the usefulness of the cell line as an in vitro model for studying trophoblast cell differentiation. A cell line was generated from choriocarcinoma explants and named Rcho-1. The cell line consisted of a mixture of cell types, including small cells growing in clusters and giant cells possessing very large nuclei. This characteristic morphology was maintained through at least 23 passages and in a series of clonal cell lines isolated from the parent Rcho-1 cell line. The Rcho-1 cell line was capable of expressing placental lactogen-I (PL-I), PL-II, PRL-like protein-A (PLP-A), and PLP-C mRNAs when cultivated in vitro; however, the Rcho-1 cells expressed only PL-I when grown beneath the kidney capsule of host rats. The Rcho-1 cell line did not express PLP-B under any experimental condition. This pattern of placental PRL expression was maintained for 23 passages. Rcho-1 cells synthesized and secreted PL-I, PL-II, and PLP-A proteins with biochemical characteristics similar to those of their placental counterparts. PL-I and PL-II mRNAs were specifically localized to giant cells. Morphological appearance and placental PRL expression were used as indices for monitoring the differentiation state of Rcho-1 cells grown under various conditions. Both morphological and functional trophoblast cell differentiation were induced by maintaining the Rcho-1 cells in postconfluent culture conditions. Postconfluent Rcho-1 cultures were characterized by an increased percentage of giant cells and an induction of placental PRL expression. Some clonal cell lines derived from the parent Rcho-1 cell line exhibited distinct patterns of differentiation and placental PRL expression. In summary, we have established a rat trophoblast cell line capable of expressing a differentiated phenotype. The differentiated phenotype includes both morphological and functional parameters and can be modulated in vitro. This cell line is a unique model for studying the control of placental PRL gene expression and the regulation of trophoblast cell differentiation.
本研究的目的是建立并鉴定一种源自大鼠绒毛膜癌的细胞系，并评估该细胞系作为研究滋养层细胞分化的体外模型的实用性。从绒毛膜癌外植体中生成了一种细胞系，并将其命名为Rcho-1。该细胞系由多种细胞类型组成，包括成簇生长的小细胞和具有非常大细胞核的巨细胞。这种特征性形态在至少23代培养以及从亲代Rcho-1细胞系分离出的一系列克隆细胞系中得以维持。Rcho-1细胞系在体外培养时能够表达胎盘催乳素-I（PL-I）、PL-II、催乳素样蛋白-A（PLP-A）和PLP-C的mRNA；然而，当Rcho-1细胞在宿主大鼠肾被膜下生长时，仅表达PL-I。在任何实验条件下，Rcho-1细胞系均不表达PLP-B。这种胎盘催乳素的表达模式在23代培养中得以维持。Rcho-1细胞合成并分泌PL-I、PL-II和PLP-A蛋白，其生化特性与其胎盘对应物相似。PL-I和PL-II的mRNA特异性定位于巨细胞。形态外观和胎盘催乳素表达被用作监测在各种条件下生长的Rcho-1细胞分化状态的指标。通过将Rcho-1细胞维持在汇合后培养条件下，诱导了形态学和功能性滋养层细胞的分化。汇合后培养的Rcho-1细胞的特征是巨细胞百分比增加以及胎盘催乳素表达的诱导。一些源自亲代Rcho-1细胞系的克隆细胞系表现出不同的分化模式和胎盘催乳素表达。总之，我们建立了一种能够表达分化表型的大鼠滋养层细胞系。分化表型包括形态学和功能参数，并且可以在体外进行调节。该细胞系是研究胎盘催乳素基因表达的控制和滋养层细胞分化调节的独特模型。
