Stadnicka Katarzyna, Marszałek Andrzej, Kozłowska Izabela, Walasik Konrad, Bodnar Magdalena, Bajek Anna, Porowińska Dorota, Drewa Tomasz, Bednarczyk Marek
Folia Biol (Krakow). 2014;62(3):179-85. doi: 10.3409/fb62_3.179.
The potential for proliferation and differentiation has a critical meaning in terms of the long-term in vitro culture of oviductal target cells. Therefore, it is important to characterize the oviduct epithelial cells, using approved markers. There is scarce data describing the epithelial cells lining the avian oviduct, most of it referring only to the magnum section of the oviduct. This study presents a comprehensive analysis of both magnum and infundibulum tissues, as the most preferred sources of epithelial cells for research on production of recombinant proteins in oviducts of birds. The main objective was to evaluate the expression of p63 and high molecular weight cytokeratins (anti- p63 antibody and anti- High Molecular Weight Cytokeratins) in epithelial cells (EC) of 2 oviduct sections: magnum (proximal and middle) and infundibulum (distal). IHC analysis and western blotting were performed using the mouse monoclonal anti- p63 antibody and anti-Ck HMW. Immunoreactivity was quantified based on the Remmele - Stegner scoring system (0-12). The expression of p63 in nuclei of luminal cells was significantly higher in the infundibulum (P < 0.05), compared to the magnum section. Cytokeratins were also highly expressed in the infundibulum, but the difference was non-significant. These findings reveal new characteristics of the oviduct EC and designate the location of the source of cells in the oviduct tissue for in vitro culture.
就输卵管靶细胞的长期体外培养而言,增殖和分化的潜力具有至关重要的意义。因此,使用经认可的标志物对输卵管上皮细胞进行表征很重要。关于禽类输卵管内衬上皮细胞的数据稀缺,其中大部分仅涉及输卵管的峡部。本研究对峡部和漏斗部组织进行了全面分析,这两个部位是禽类输卵管中用于重组蛋白生产研究的上皮细胞的最优选来源。主要目的是评估p63和高分子量细胞角蛋白(抗p63抗体和抗高分子量细胞角蛋白)在两个输卵管切片的上皮细胞(EC)中的表达:峡部(近端和中部)和漏斗部(远端)。使用小鼠单克隆抗p63抗体和抗Ck HMW进行免疫组化分析和蛋白质印迹。基于Remmele - Stegner评分系统(0 - 12)对免疫反应性进行定量。与峡部相比,漏斗部管腔细胞核中p63的表达显著更高(P < 0.05)。细胞角蛋白在漏斗部也高度表达,但差异不显著。这些发现揭示了输卵管EC的新特征,并确定了用于体外培养的输卵管组织中细胞来源的位置。