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Optimizing culture conditions of a porcine epithelial cell line IPEC-J2 through a histological and physiological characterization

机译:通过组织学和生理学表征优化猪上皮细胞系IPEC-J2的培养条件

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摘要

The high similarity between pigs and humans makes pigs a good gastrointestinal (GI) model for humans. Recently an epithelial cell line originating from the jejunum of pig (IPEC-J2) became available. Once validated, this model can be used to investigate the complex interactions occurring in the intestine. The advantages of using IPEC-J2 as in vitro model of the GI tract are the high resemblance between humans and pigs, and the ease of extrapolating in vitro to in vivo characteristics. In this study, the IPEC-J2 cells were functionally characterized by measuring the trans-epithelial electrical resistance (TEER), and by histological and ultrastructural studies. IPEC-J2 cells grown on six different permeable support systems, were investigated. The Transwell®-COL collagen-coated membrane (1.12 cm2) showed the best results concerning time efficiency and TEER values. The optimum seeding density of 12 × 105 cells/mL ensured that after 9 days of differentiation a confluent monolayer was formed. The decrease in TEER values after a maximum had been reached, coincided with the ultrastructural development of apical microvilli. We conclude that IPEC-J2 cells grown on collagen-coated membranes represent a valuable in vitro model system for the small intestinal epithelium which can be of great interest for intestinal research.
机译:猪与人之间的高度相似性使猪成为人的良好胃肠道(GI)模型。最近,可获得来自猪空肠的上皮细胞系(IPEC-J2)。一旦验证,该模型可用于研究肠道中发生的复杂相互作用。使用IPEC-J2作为胃肠道的体外模型的优点是人与猪之间的高度相似性,以及易于从体外推断到体内的特征。在这项研究中,通过测量跨上皮电阻(TEER)以及组织学和超微结构研究对IPEC-J2细胞进行了功能表征。研究了在六个不同的渗透性支持系统上生长的IPEC-J2细胞。 Transwell ® -COL胶原蛋白涂层膜(1.12cm 2 )在时间效率和TEER值方面显示出最好的结果。最佳接种密度为12×10 5 细胞/ mL,可确保分化9天后形成汇合的单层细胞。达到最大值后,TEER值下降,与顶端微绒毛的超微结构发展相吻合。我们得出的结论是,在胶原蛋白包被的膜上生长的IPEC-J2细胞代表了小肠上皮细胞的有价值的体外模型系统,这对肠道研究可能非常感兴趣。

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