Choudhury Jaganmoy, Richardson Lauren S, Urrabaz-Garza Rheanna, Jacob Jeena, Kammala Ananth Kumar, Menon Ramkumar
Division of Basic Science and Translational Research, Department of Obstetrics and Gynecology, The University of Texas Medical Branch at Galveston, Galveston, TX, United States of America.
Biol Reprod. 2025 Mar 16;112(3):530-539. doi: 10.1093/biolre/ioaf003.
Chorionic trophoblast cells are one of the principal components of the fetal membrane and join with the decidua to form a feto-maternal interface. Recent success in isolating chorionic trophoblast cells dealt with two separate questions: (i) the necessity of highly enriched and defined media with inhibitors of oxidative stress and cell transition and their impact on growth and trophoblast phenotype, (ii) the functional differences between chorionic trophoblast cells and other placental trophoblast lineages of cells (placental cytotrophoblast cells, and extravillous trophoblast). Chorionic trophoblast cells were cultured either in defined media with various inhibitors or in media from which inhibitors were removed individually. Cellular morphology and growth (microscopy and crystal violet staining) and cellular and molecular biological features (immunofluorescence staining for GATA-binding protein 3, cytokeratin 7, and vimentin) were assessed. Syncytialization of cells (forskolin treatment) and invasive properties of chorionic trophoblast cells (cell invasion assay) were tested and compared with placental cytotrophoblast cells and extravillous trophoblasts (HTR8/SVneo), respectively. Removal of various growth-supporting agents from the media delayed cell growth and inclined towards cellular transition (increase in vimentin compared to cytokeratin 7 or GATA-binding protein 3) compared to chorionic trophoblast cells grown in complete and enriched media. The chorionic trophoblast cells failed to syncytialize, contrasting with the high levels of membrane fusion observed in placental cytotrophoblast cells. Although chorionic trophoblast cells express human leukocyte antigen G like extravillous trophoblasts, they do not invade. Chorionic trophoblast cells require several specific constituents for in vitro growth and phenotype maintenance. Chorionic trophoblast cells are trophoblast lineage cells that barricade immune cell-enriched decidua without invading them. These properties support their location and function, which are distinct from placental cytotrophoblast cells and extravillous trophoblasts.
绒毛膜滋养层细胞是胎膜的主要成分之一,与蜕膜共同形成母胎界面。近期在分离绒毛膜滋养层细胞方面取得的成功解决了两个不同的问题:(i)含有氧化应激抑制剂和细胞转化抑制剂的高度富集且明确的培养基的必要性及其对生长和滋养层细胞表型的影响,(ii)绒毛膜滋养层细胞与其他胎盘滋养层细胞谱系(胎盘细胞滋养层细胞和绒毛外滋养层细胞)之间的功能差异。绒毛膜滋养层细胞分别在含有各种抑制剂的明确培养基中或在逐个去除抑制剂的培养基中培养。评估细胞形态和生长(显微镜检查和结晶紫染色)以及细胞和分子生物学特征(针对GATA结合蛋白3、细胞角蛋白7和波形蛋白的免疫荧光染色)。测试细胞的合体化(福斯高林处理)和绒毛膜滋养层细胞的侵袭特性(细胞侵袭试验),并分别与胎盘细胞滋养层细胞和绒毛外滋养层细胞(HTR8/SVneo)进行比较。与在完全富集培养基中生长的绒毛膜滋养层细胞相比,从培养基中去除各种生长支持剂会延迟细胞生长并倾向于细胞转化(与细胞角蛋白7或GATA结合蛋白3相比,波形蛋白增加)。绒毛膜滋养层细胞未能合体化,这与在胎盘细胞滋养层细胞中观察到的高水平膜融合形成对比。尽管绒毛膜滋养层细胞像绒毛外滋养层细胞一样表达人类白细胞抗原G,但它们并不侵袭。绒毛膜滋养层细胞在体外生长和表型维持需要几种特定成分。绒毛膜滋养层细胞是滋养层细胞谱系细胞,它们隔离富含免疫细胞的蜕膜而不侵入其中。这些特性支持了它们与胎盘细胞滋养层细胞和绒毛外滋养层细胞不同的定位和功能。