Laboratory of Reproductive Physiology, Graduate School of Environmental and Life Science, Okayama University, Okayama 700-8530, Japan.
Laboratory of Reproductive Physiology, Faculty of Agriculture, Okayama University, Okayama 700-8530, Japan.
J Reprod Dev. 2022 Aug 1;68(4):254-261. doi: 10.1262/jrd.2022-029. Epub 2022 May 27.
In ruminants, uterine glands play key roles in the establishment of pregnancy by secreting various factors into the uterine lumen. Although a three-dimensional (3D) culture system has been used for investigating cellular functions in vitro, the detailed functions of uterine gland have not been fully elucidated. In this study, we examined the benefits of 3D culture system to examine the innate functions of bovine uterine glands. Isolated bovine uterine glands were cultured on Matrigel (2D) or in Matrigel (3D), respectively, and the mRNA levels of secreted proteins (SERPINA14, MEP1B, APOA1, ARSA, CTGF, and SPP1) were measured in isolated and cultured uterine glands. The protein expression of estrogen receptor β (ERβ) and progesterone receptor (PR) and the establishment of apico-basal polarity were examined. In isolated uterine glands, the mRNA levels of secreted proteins changed during the estrous cycle. Although uterine glands cultured in both 2D and 3D expressed ERβ and PR, progesterone did not affect SERPINA14 mRNA expression. The expression of APOA1 mRNA in 2D cultured uterine glands did not respond to estrogen and progesterone. Additionally, the mRNA levels of secreted proteins in the 3D culture system were significantly higher than those in the 2D culture system, which might be attributed to the different cellular morphology between them. The locations of ZO-1 and β-catenin in 2D cultured uterine glands were disordered compared with 3D cultured uterine glands. These results showed that the hormonal responsiveness of secreted factor expression and cellular morphology were different between 2D and 3D cultured bovine uterine glands.
在反刍动物中，子宫腺通过将各种因子分泌到子宫腔中，在妊娠的建立中发挥关键作用。虽然已经使用三维（3D）培养系统来研究体外细胞功能，但子宫腺的详细功能尚未完全阐明。在这项研究中，我们检查了 3D 培养系统在检查牛子宫腺固有功能方面的好处。分别将分离的牛子宫腺在 Matrigel（2D）或 Matrigel（3D）上培养，并测量分离和培养的子宫腺中分泌蛋白（SERPINA14、MEP1B、APOA1、ARSA、CTGF 和 SPP1）的 mRNA 水平。检查了雌激素受体β（ERβ）和孕激素受体（PR）的蛋白表达以及顶-基底极性的建立。在分离的子宫腺中，分泌蛋白的 mRNA 水平在发情周期中发生变化。尽管在 2D 和 3D 培养的子宫腺中均表达 ERβ和 PR，但孕激素不影响 SERPINA14 mRNA 的表达。在 2D 培养的子宫腺中，APOA1 mRNA 的表达对雌激素和孕激素没有反应。此外，3D 培养系统中分泌蛋白的 mRNA 水平明显高于 2D 培养系统，这可能归因于它们之间不同的细胞形态。与 3D 培养的子宫腺相比，2D 培养的子宫腺中 ZO-1 和β-连环蛋白的位置紊乱。这些结果表明，2D 和 3D 培养的牛子宫腺中分泌因子表达的激素反应性和细胞形态不同。
