Co-culture cassette for anaerobes and primary human intestinal epithelium

项目摘要 人类结肠是一个显著的器官，在药物摄取和代谢中起着关键作用。 代谢以及窝藏微生物组的100万亿个微生物细胞， 对人类健康有多重影响。由于这些原因，广泛需要 人类结肠生理学体外研究的学术界和生物技术市场 以及结肠组织和微生物组中的厌氧菌之间的相互作用。满足 Altis Biosystems，Inc.一家早期的生物技术公司，与 位于查佩尔山的北卡罗来纳州大学的科学家们开发了一个新的平台 对于细胞培养，将正常的人结肠上皮细胞与厌氧微生物群共培养， 自我维持肠道微生物组平台（SIMPle）。SIMPle是一个易于使用的 和直观的平台来复制跨越体内结肠的陡峭氧气梯度 上皮，从而创造适当的环境所需的厌氧菌，同时保持 健康的上皮组织我们已经完成了SBIR第一阶段计划的设计， 原型设计，验证适合标准12孔板的SIMPLe（SIMPle 12）。生成 的陡峭的O2梯度，并确定其对细胞的影响。共培养 在SIMPle 12平台中，对兼性厌氧菌与人结肠上皮的分离进行了验证。 第一阶段SBIR中的所有拟议里程碑都已完成，从而提供了坚实的 这是第二阶段SBIR应用的基础。第二阶段提案的重点是扩大 SIMPLe平台升级为96孔格式（SIMPle 96），以满足市场对筛选分析的需求。 SIMPle 96平台将进行原型设计和验证，人类结肠上皮和 将表征与专性厌氧菌的共培养。使用SIMPle 96平台进行筛选 益生菌将被证明。将对平台进行外部确认， 证明了其在微生物-肠道测定中的实用性。SIMPle共培养平台包括 有望通过使宿主微生物的功能， 互动要审问。

Project Summary The human colon is a remarkable organ, playing critical roles in drug uptake and metabolism as well as harboring the 100 trillion microbial cells of the microbiome, which itself has multiple impacts on human health. For these reasons, there is a widespread need in academia and the biotechnology marketplace for in vitro studies of human colon physiology and the interaction between colon tissue and the anaerobic bacteria of the microbiome. To meet this need, Altis Biosystems, Inc., an early stage biotechnology company, has collaborated with scientists at the University of North Carolina (UNC) at Chapel Hill to develop a new platform for cell culture to co-culture normal, human colonic epithelial cells with anaerobic microbiota, named the Self-sustaining Intestinal Microbiome Platform (SIMPle). SIMPle is an easy-to-use and intuitive platform to replicate the steep oxygen gradient across the in vivo colonic epithelium, thus create the appropriate environment required for anaerobes while maintaining viable, healthy epithelial tissue. We have finished the SBIR Phase I program by designing, prototyping, validating the SIMPLe that fits a standard 12-well plate (SIMPle12). The generation of a steep O2 gradient was confirmed, and its impact on cells was characterized. The co-culture of facultative anaerobes with human colonic epithelium in SIMPle12 platforms was validated. All proposed milestones in the Phase I SBIR were accomplished, thus providing a solid foundation for this Phase II SBIR application. The focus of this Phase II proposal is to scale up SIMPLe platform to a 96-well format (SIMPle96) to meet the market needs for screening assays. The SIMPle96 platform will be prototyped and validated, and human colonic epithelium and co-culture with obligate anaerobes will be characterized. Use SIMPle96 platform for screening probiotics will be demonstrated. An external validation of the platform will be performed to demonstrate its utility in microbe-intestinal assays. The SIMPle co-culture platforms are expected to revolutionize microbiome research by enabling the function of host-microbe interactions to be interrogated.