Kim Raehyun, Wang Yuli, Sims Christopher E, Allbritton Nancy L
Department of Biological and Chemical Engineering, Hongik University, Sejong, South Korea.
Department of Bioengineering, University of Washington, Seattle, WA, United States.
Front Bioeng Biotechnol. 2022 Jun 8;10:890396. doi: 10.3389/fbioe.2022.890396. eCollection 2022.
An platform was designed and optimized for the co-culture of probiotic anaerobic bacteria with a primary human colonic epithelium having a goal of assessing the anti-inflammatory impact of the probiotic bacteria. The device maintained a luminal O concentration at <1% while also supporting an oxygenated basal compartment at 10% for at least 72 h. Measurement of the transepithelial resistance of a confluent colonic epithelium showed high monolayer integrity while fluorescence assays demonstrated that the monolayer was comprised primarily of goblet cells and colonocytes, the two major differentiated cell subtypes of the colonic epithelium. High monolayer barrier function and viability were maintained during co-culture of the epithelium with the probiotic obligate anaerobe (). Importantly the device supported a static co-culture of microbes and colonic epithelium mimicking the largely static or low flow conditions within the colonic lumen. A model inflamed colonic epithelium was generated by the addition of tumor necrosis factor-α (TNF-α) and lipopolysaccharide (LPS) to the basal and luminal epithelium sides, respectively. Co-culture of with the LPS/TNF-α treated intestine diminished IL-8 secretion by ≥40% which could be mimicked by co-culture with the metabolite butyrate. In contrast, co-culture of the inflamed epithelium with two strains of lactic acid-producing bacteria, () and (), did not diminish epithelial IL-8 secretion. Co-culture with colonic epithelial cells from different donors demonstrated a consistent anti-inflammatory effect by , but distinct responses to co-culture with and . The demonstrated system offers a simple and easily adopted platform for examining the physiologic impact of alterations in the intestinal epithelium that occur in the presence of probiotic bacteria and their metabolites.
设计并优化了一个平台，用于益生菌厌氧菌与原代人结肠上皮细胞的共培养，目的是评估益生菌的抗炎作用。该装置将管腔氧浓度维持在<1%，同时在至少72小时内将含氧量为10%的基底隔室维持在该水平。对汇合的结肠上皮细胞的跨上皮电阻测量显示单层完整性高，而荧光分析表明该单层主要由杯状细胞和结肠细胞组成，这是结肠上皮的两种主要分化细胞亚型。在上皮细胞与专性厌氧益生菌（）共培养期间，维持了高单层屏障功能和活力。重要的是，该装置支持微生物与结肠上皮细胞的静态共培养，模拟结肠腔内很大程度上的静态或低流量条件。通过分别在基底和管腔上皮侧添加肿瘤坏死因子-α（TNF-α）和脂多糖（LPS），生成了模型性炎症结肠上皮。与经LPS/TNF-α处理的肠道共培养可使IL-8分泌减少≥40%，这可通过与代谢产物丁酸盐共培养来模拟。相比之下，炎症上皮与两株产乳酸菌（）和（）共培养并未减少上皮IL-8分泌。与来自不同供体的结肠上皮细胞共培养显示了一致的抗炎作用，但对与和共培养的反应不同。所展示的系统为研究在存在益生菌及其代谢产物的情况下肠道上皮细胞变化的生理影响提供了一个简单且易于采用的平台。
