牛子宫腺在二维和三维培养系统中的功能评价

Yosuke Sugino, Taiki Sato, Yuki Yamamoto, K. Kimura
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引用次数: 1

摘要

在反刍动物中,子宫腺通过将各种因子分泌到子宫腔中,在妊娠的建立中起着关键作用。虽然三维(3D)培养系统已被用于体外研究细胞功能,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培养系统检查牛子宫腺先天功能的好处。分别在Matrigel (2D)和Matrigel (3D)上培养离体牛子宫腺,测定离体和培养的子宫腺中分泌蛋白(SERPINA14、MEP1B、APOA1、ARSA、CTGF、SPP1) mRNA水平。研究了雌激素受体β (ERβ)和孕激素受体(PR)的蛋白表达及尖基极性的建立。在离体子宫腺中,分泌蛋白mRNA水平在发情周期中发生变化。2D和3D子宫腺均表达ERβ和PR,但黄体酮不影响SERPINA14 mRNA的表达。APOA1 mRNA在2D培养子宫腺中的表达对雌激素和黄体酮无反应。此外,3D培养系统分泌蛋白mRNA水平明显高于2D培养系统,这可能与两者细胞形态不同有关。与3D培养子宫腺相比,ZO-1和β-catenin在2D培养子宫腺中的位置紊乱。结果表明,二维和三维培养的牛子宫腺分泌因子表达和细胞形态的激素反应性存在差异。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Evaluation of bovine uterine gland functions in 2D and 3D culture system
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.
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