Liu Bingying, Ren Siqi, An Hong, Liang Yixuan, Sheng Xihui, Qi Xiaolong, Xiao Longfei, Wang Xiangguo
College of Bioscience and Resources Environment, Beijing University of Agriculture, Beijing 102206, China.
Animal Science and Technology College, Beijing University of Agriculture, Beijing 102206, China.
Cells Dev. 2024 Dec;180:203970. doi: 10.1016/j.cdev.2024.203970. Epub 2024 Sep 5.
The placenta is an organ that plays a vital role in successful pregnancies, and the failure of early placentation is a significant factor leading to abortion in ruminant species. However, the mechanisms involved in the development and differentiation of bovine placenta remain elusive due to the lack of suitable in vitro placental models. This study aimed to develop an effective method for generating the bovine functional trophoblast organoids by assembling bovine primary trophoblast cells (PBTCs) from the placenta or immortalized bovine placental trophoblast (BTCs) in a 3D culture system in vitro. PBTCs isolated from the 3-month-gestation placenta and BTCs rapidly proliferated and exhibited typical epithelioid morphology in the modified trophoblast organoid medium (TOM) for bovine. Furthermore, PBTCs and BTCs proliferating in the modified TOM were both CK7- and E-cadherin-positive. Both PBTCs or BTCs embedded into Matrigel droplets overlaid with modified TOM proliferated and formed trophoblast organoids after 15 days of culture. Moreover, the expression of syntrophoblast marker genes, including CD71, CD46, and chorionic somatomammotropin hormone 1 (CSH1), was detectable in both organoids derived from different types of trophoblast cells. Notably, the protein expression levels of various genes implicated in the establishment of early pregnancy in endometrial epithelium cells (EECs) was increased following coculture with bovine trophoblast organoids. Collectively, the bovine trophoblast organoids established in our study could serve as robust models for elucidating the essential physical functions of the placenta and the causes of pregnancy failures related to the placenta developmental disorders during early bovine pregnancy.
胎盘是在成功妊娠中发挥至关重要作用的器官，早期胎盘形成失败是反刍动物流产的重要因素。然而，由于缺乏合适的体外胎盘模型，牛胎盘发育和分化所涉及的机制仍不清楚。本研究旨在开发一种有效的方法，通过在体外三维培养系统中组装来自胎盘的牛原代滋养层细胞（PBTCs）或永生化牛胎盘滋养层细胞（BTCs）来生成具有功能的牛滋养层类器官。从妊娠3个月的胎盘分离出的PBTCs和BTCs在改良的牛滋养层类器官培养基（TOM）中迅速增殖，并呈现典型的上皮样形态。此外，在改良的TOM中增殖的PBTCs和BTCs均为CK7和E-钙黏蛋白阳性。将PBTCs或BTCs嵌入覆盖有改良TOM的基质胶滴中，培养15天后均增殖并形成滋养层类器官。此外，在源自不同类型滋养层细胞的两种类器官中均检测到合体滋养层标记基因的表达，包括CD71、CD46和绒毛膜生长催乳素1（CSH1）。值得注意的是，与牛滋养层类器官共培养后，子宫内膜上皮细胞（EECs）中与早期妊娠建立相关的各种基因的蛋白表达水平有所增加。总体而言，我们研究中建立的牛滋养层类器官可作为强大的模型，用于阐明胎盘的基本生理功能以及早期牛妊娠期间与胎盘发育障碍相关的妊娠失败原因。
