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利用新型酶基分离方法区分小鼠子宫腔上皮和腺体上皮的特征。

Distinguish Characters of Luminal and Glandular Epithelium from Mouse Uterus Using a Novel Enzyme-Based Separation Method.

机构信息

College of Veterinary Medicine, South China Agricultural University, 483 Wushan Road, Guangzhou, Guangdong, 510642, China.

出版信息

Reprod Sci. 2023 Jun;30(6):1867-1877. doi: 10.1007/s43032-022-01154-z. Epub 2022 Dec 29.

Abstract

The uterine luminal epithelium, glandular epithelium, and stromal cells are vital for the establishment of pregnancy. Previously studies have shown various methods to isolate mouse uterine epithelium and stromal cells, including laser capture microdissection (LCM), enzyme digestion, and immunomagnetic beads. Despite the importance of the endometrial epithelium as the site of implantation and nutritional support for the conceptus, there is no isolated method to separate the luminal epithelium and glandular epithelium. Here, we establish a novel enzyme-based way to separate two types of epithelium and keep their viability. In this article, we analyzed their purity by mRNA level, immunostaining, and transcriptome analysis. Our isolation method revealed several unstudied luminal and glandular epithelial markers in transcriptome analysis. We further demonstrated the viability of the isolated epithelium by 2D and 3D cultures. The results showed that we successfully separated the endometrial luminal epithelium and glandular epithelium. We also provided an experimental model for the following study of the physiological function of the different parts of the uterus and related diseases.

摘要

子宫腔上皮、腺上皮和基质细胞对于妊娠的建立至关重要。先前的研究已经提出了各种分离小鼠子宫上皮和基质细胞的方法,包括激光捕获显微切割(LCM)、酶消化和免疫磁珠。尽管子宫内膜上皮作为着床部位和对胚胎的营养支持非常重要,但目前还没有分离腔上皮和腺上皮的方法。在这里,我们建立了一种新型的基于酶的方法来分离两种上皮细胞,并保持其活力。在本文中,我们通过 mRNA 水平、免疫染色和转录组分析来分析它们的纯度。我们的分离方法在转录组分析中揭示了一些未被研究的腔上皮和腺上皮标志物。我们通过 2D 和 3D 培养进一步证明了分离的上皮细胞的活力。结果表明,我们成功地分离了子宫内膜腔上皮和腺上皮。我们还为研究子宫不同部位的生理功能和相关疾病提供了一个实验模型。

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