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Identification of pluripotent cells in bovine uterus: in situ and in vitro studies

机译:牛子宫多能细胞的鉴定:原位和体外研究

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The aim of this study was to identify uterine pluripotent cells both in bovine uterine tissues as well in epithelial, stromal, and myometrial uterine cell populations. Moreover, the relationship of pluripotent markers expression with age and the uterine horn side was considered. Uterine tissue was collected from ipsilateral and contralateral horns (days 8–10 of the estrous cycle). Immunohistostaining for C-KIT, OCT3/4, NANOG, and SOX2 in uterine tissue was determined. mRNA expression of C-KIT, OCT3/4, NANOG and SOX2 was evaluated in uterine tissue relative to the age of the cow and uterine horn side. Gene and protein expression of these markers in the uterine luminal epithelial, stromal, and myometrial cells was evaluated by real-time PCR and western blotting respectively. The expression of pluripotent cell markers OCT3/4, NANOG, and SOX2 was identified by flow cytometry assay in epithelial, stromal, and myometrial cells. Multilineage differentiation of the bovine uterine cells was performed. mRNA expression of OCT3/4, NANOG, and SOX2 in uterine tissue was higher in the ipsilateral horn than in the contralateral horn. Flow cytometry assay revealed positive fluorescence for OCT3/4, NANOG, and SOX2 in all uterine cell types. Results showed the age-dependent expression of pluripotent markers in uterine tissue. Beside, the different expression of pluripotent cells in each horn of uterus suggests the influence of ovarian hormones on these characteristics. The highest mRNA and protein expression for pluripotent markers was observed in stromal cells among uterine cells, which indicates this population of cells as the main site of pluripotent cells in the cow uterus.
机译:这项研究的目的是在牛子宫组织以及上皮,基质和子宫肌层子宫细胞群中鉴定子宫多能细胞。此外,考虑了多能标志物表达与年龄和子宫角侧的关系。从同侧和对侧角(动情周期的第8-10天)收集子宫组织。确定了子宫组织中C-KIT,OCT3 / 4,NANOG和SOX2的免疫组织染色。相对于母牛和子宫角侧的年龄,评估了子宫组织中C-KIT,OCT3 / 4,NANOG和SOX2的mRNA表达。分别通过实时PCR和蛋白质印迹法评估这些标志物在子宫腔上皮细胞,基质细胞和子宫肌层细胞中的基因和蛋白质表达。通过流式细胞术测定在上皮,基质和肌层细胞中鉴定多能细胞标志物OCT3 / 4,NANOG和SOX2的表达。进行了牛子宫细胞的多谱系分化。同侧角的子宫组织中OCT3 / 4,NANOG和SOX2的mRNA表达高于对侧角。流式细胞仪检测显示所有类型的子宫细胞中OCT3 / 4,NANOG和SOX2均呈阳性荧光。结果显示子宫组织中多能性标记物的年龄依赖性表达。此外,子宫各个角中多能细胞的不同表达提示卵巢激素对这些特征的影响。在子宫细胞中的间质细胞中观察到多能标记的最高mRNA和蛋白质表达,这表明该细胞群是牛子宫中多能细胞的主要位点。

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