Department of Reproduction Biology, Leibniz-Institute for Zoo and Wildlife Research (IZW), Alfred-Kowalke-Straße 17, 10315, Berlin, Germany.
Institute of Reproductive Biology, Research Institute for Farm Animal Biology (FBN), 18196, Dummerstorf, Germany.
Histochem Cell Biol. 2024 Jun;161(6):521-537. doi: 10.1007/s00418-024-02273-1. Epub 2024 Mar 26.
Key reproductive events such as fertilization and early embryonic development occur in the lumen of the oviduct. Since investigating these processes in vivo is both technically challenging and ethically sensitive, cell culture models have been established to reproduce the oviductal microenvironment. Compartmentalized culture systems, particularly air-liquid interface cultures (ALI; cells access the culture medium only from the basolateral cell side), result in highly differentiated oviduct epithelial cell cultures. The oxygen (O) tension within the oviduct is 4-10% across species, and its reduced O content is presumed to be important for early reproductive processes. However, cell culture models of the oviduct are typically cultivated without O regulation and therefore at about 18% O. To investigate the impact of O levels on oviduct epithelium functions in vitro, we cultured porcine oviduct epithelial cells (POEC) at the ALI using both physiological (5%) and supraphysiological (18%) O levels and two different media regimes. Epithelium architecture, barrier function, secretion of oviduct fluid surrogate (OFS), and marker gene expression were comparatively assessed. Under all culture conditions, ALI-POEC formed polarized, ciliated monolayers with appropriate barrier function. Exposure to 18% O accelerated epithelial differentiation and significantly increased the apical OFS volume and total protein content. Expression of oviduct genes and the abundance of OVGP1 (oviduct-specific glycoprotein 1) in the OFS were influenced by both O tension and medium choice. In conclusion, oviduct epithelial cells can adapt to a supraphysiological O environment. This adaptation, however, may alter their capability to replicate in vivo tissue characteristics.
受精和早期胚胎发育等关键生殖事件发生在输卵管的管腔中。由于在体内研究这些过程既具有技术挑战性，又涉及伦理敏感性，因此建立了细胞培养模型来复制输卵管的微环境。分隔培养系统，特别是气液界面培养（ALI；细胞只能从基底外侧细胞侧进入培养基），导致高度分化的输卵管上皮细胞培养。输卵管内的氧（O）张力在各个物种之间为 4-10%，其减少的 O 含量被认为对早期生殖过程很重要。然而，输卵管的细胞培养模型通常在没有 O 调节的情况下进行培养，因此 O 含量约为 18%。为了研究 O 水平对体外输卵管上皮功能的影响，我们使用生理（5%）和超生理（18%）O 水平以及两种不同的培养基方案在 ALI 上培养猪输卵管上皮细胞（POEC）。比较评估了上皮结构、屏障功能、输卵管液替代物（OFS）的分泌和标记基因表达。在所有培养条件下，ALI-POEC 形成极化的、纤毛单层，具有适当的屏障功能。暴露于 18%O 加速了上皮分化，并显著增加了顶端 OFS 体积和总蛋白含量。输卵管基因的表达和 OFS 中 OVGP1（输卵管特异性糖蛋白 1）的丰度受 O 张力和培养基选择的影响。总之，输卵管上皮细胞可以适应超生理的 O 环境。然而，这种适应可能会改变它们在体内复制组织特征的能力。
