PDF(Pubmed)
Abstract：
The complexification of in vitro models requires the compatibility of cells with the same medium. Since immune cells are the most sensitive to growth conditions, growing intestinal epithelial cells in their usual medium seems to be necessary. This work was aimed at comparing the sensitivity of these epithelial cells to pro-inflammatory stimuli but also to dietary polyphenols in both DMEM and RPMI-1640 media. Co-cultures of Caco-2 and HT29-MTX cells were grown for 21 days in the two media before their stimulation with a cocktail of TNF-α (20 ng/mL), IL-1β (1 ng/mL), and IFN-γ (10 ng/mL) or with LPS (10 ng/mL) from E. coli (O111:B4). The role of catechins (15 µM), a dietary polyphenol, was evaluated after its incubation with the cells before their stimulation for 6 h. The RPMI-1640 medium did not alter the intensity of the inflammatory response observed with the cytokines. By contrast, LPS failed to stimulate the co-culture in inserts regardless of the medium used. Lastly, catechins were unable to prevent the pro-inflammatory response observed with the cytokines in the two media. The preservation of the response of this model of intestinal epithelium in RPMI-1640 medium is promising when considering its complexification to evaluate the complex cellular crosstalk leading to intestinal homeostasis.
摘要：
体外模型的复合需要细胞与相同培养基的相容性。由于免疫细胞对生长条件最敏感，在通常的培养基中生长肠上皮细胞似乎是必要的。这项工作旨在比较这些上皮细胞对促炎刺激的敏感性，以及对DMEM和RPMI-1640培养基中膳食多酚的敏感性。Caco-2和HT29-MTX细胞的共培养物在两种培养基中生长21天，然后用TNF-α（20ng/mL）的混合物进行刺激，IL-1β（1ng/mL），和IFN-γ(10ng/mL)或来自大肠杆菌(O111:B4)的LPS(10ng/mL)。儿茶素（15µM）的作用，一种膳食多酚，在其刺激之前与细胞孵育6小时后进行评估。RPMI-1640培养基没有改变用细胞因子观察到的炎症反应的强度。相比之下,无论使用何种培养基,LPS都不能刺激插入物中的共培养物。最后,儿茶素不能预防两种培养基中细胞因子的促炎反应.当考虑其复合性以评估导致肠稳态的复杂细胞串扰时，在RPMI-1640培养基中保留该肠上皮模型的响应是有希望的。
