A Novel Method to Improve Function of SC-Derived Hepatocytes

描述（由申请人提供）：Primorigen Biosciences SBIR提案摘要Primorigen Biosciences将使用SBIR资金开发先进的细胞培养方法，以提高定向分化的效率，并促进诱导多能干细胞衍生肝细胞的功能成熟。这项新技术使用专有的细胞培养基添加剂来重建细胞微环境的天然生物物理特性，并鼓励细胞分泌和重塑细胞外基质，使用其自身的内源性机制来促进祖细胞增殖和更完全的分化，以类似于原代肝细胞功能。I期研究将确定在分化和成熟过程中应用新技术的最佳参数，使用已建立的肝细胞表征试验评估分化肝细胞的功能和成熟度。II期研究将验证和优化化合物代谢和II期酶的活性，并使改进的肝细胞分化方案适应更高通量的形式，如96和384孔板，以实现药物发现和毒性研究的化合物筛选。该产品将通过Primorigen的直接客户群和主要战略联盟合作伙伴在全球范围内进行商业化。 公共卫生相关性：Primorigen Biosciences，Inc. SBIR项目叙述世界范围内缺乏用于治疗和研究应用的原代肝细胞，包括药物发现和毒性筛选。此外，原代肝细胞在组织培养环境中丧失大部分肝功能的趋势表明非常需要已知基因型的肝细胞样细胞的可扩展和可再现来源。虽然目前的干细胞培养方法在产生模拟原代肝细胞的几种功能的肝细胞样细胞方面是成功和有效的，但它们在产生完全模拟原代细胞功能的细胞方面还没有成功，特别是编码对毒性分析和关键肝功能至关重要的p450细胞色素酶的基因的表达。SBIR技术将使用专有的、正在申请专利的培养基添加剂来解决这一问题，这些添加剂使干细胞能够利用其自身的内源性机制创造一个更类似于体内的微环境，以产生功能更全的干细胞衍生的肝细胞。

DESCRIPTION (provided by applicant): Primorigen Biosciences SBIR Proposal Abstract Primorigen Biosciences will use SBIR funds to develop an advanced cell culturing methodology to increase the efficiency of directed differentiation and to boost functional maturity of induced pluripotent stem cell-derived hepatocytes. The new technology uses proprietary cell culture media additives to recreate native biophysical properties of the cellular microenvironment and encourage cells to secrete and remodel extracellular matrix using their own endogenous machinery to boost progenitor cell proliferation and more complete differentiation to resemble primary hepatocyte function. Phase I studies will determine the optimal parameters for applying the new technology during differentiation and maturation, using established hepatocyte characterization assays to assess the functionality and maturity of the differentiated hepatocytes. Phase II studies will verify and optimize compound metabolism and activity of Phase II enzymes, and adapt the improved hepatocyte differentiation protocol to higher-throughput formats such as 96 and 384 well plates to enable compound screening for drug discovery and toxicity studies. The product will be commercialized globally through Primorigen's direct customer base and major strategic alliance partners. PUBLIC HEALTH RELEVANCE: Primorigen Biosciences, Inc. SBIR Project Narrative There is a worldwide shortage of primary hepatocytes for use in both therapeutic and research applications, including drug discovery and toxicity screening. In addition, the tendency of primary hepatocytes to lose most hepatic functions in a tissue culture environment suggests a great need for a scalable and reproducible source of hepatocyte-like cells of known genotype. While current stem cell culturing methods have been successful and efficient in generating hepatocyte-like cells that mimic several functions of primary liver cells, they have not been successful in producing cells that completely mimic primary cell function, particularly expression of genes encoding for p450 cytochrome enzymes critical for toxicity analysis and key liver functions. The SBIR technology will address this problem using proprietary, patent-pending media additives that enable stem cells to create a more in vivo-like microenvironment using their own endogenous mechanisms to produce a more fully functional stem cell-derived hepatocyte.