Co-culture cassette for anaerobes and primary human intestinal epithelium

项目摘要 人的结肠是一个非凡的器官，在药物摄取和 以及拥有100万亿微生物组的微生物细胞，而微生物组本身 对人类健康有多种影响。出于这些原因，人们普遍需要 人体结肠生理学体外研究的学术界和生物技术市场 以及结肠组织与微生物群中的厌氧细菌之间的相互作用。相遇 在这一需求下，早期生物技术公司Altis BiosSystems，Inc.与 北卡罗来纳大学(UNC)教堂山分校的科学家开发一种新平台 为了让细胞培养将正常的人结肠上皮细胞与厌氧微生物区系共培养， 命名为自给自足的肠道微生物组平台(SIMPLE)。Simple是一种易于使用的 和直观的平台来复制体内结肠中陡峭的氧气梯度 上皮，从而为厌氧菌创造适当的环境，同时保持 活的、健康的上皮组织。我们已经完成了SBIR一期项目的设计， 原型制作，验证适合标准12孔板的SIMPLE(SIMPle12)。这一代人 证实了陡峭的氧气梯度的存在，并表征了其对细胞的影响。共同文化 在SIMPle12平台上验证了兼性厌氧菌与人结肠上皮的关系。 第一阶段SBIR的所有拟议里程碑都已完成，从而提供了坚实的 为第二阶段SBIR应用奠定基础。此第二阶段计划的重点是扩大规模 Simple Platform to a 96-Well Format(SIMPle96)以满足市场对筛查分析的需求。 SIMPle96平台将被原型和验证，人类结肠上皮和 将描述与专性厌氧菌共培养的特征。使用SIMPle96平台进行筛选 将演示益生菌。将对平台进行外部验证，以 展示其在微生物肠道检测中的有效性。简单的共同文化平台是 有望通过实现宿主-微生物的功能来彻底改变微生物组的研究 相互作用有待审问。

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.