A Novel Method to Improve Function of SC-Derived Hepatocytes

改善 SC 源性肝细胞功能的新方法

• 批准号: 8314618
• 负责人: Scott Allen Monsma
• 金额: $ 32.82万
• 依托单位: PRIMORIGEN BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-01 至 2013-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8314618
• 关键词: Albumins; Biological Assay; Carbohydrates; Categories; Cell Culture Techniques; Cell Differentiation process; Cell Proliferation; Cell Therapy; Cells; Collagen; Crowding; Culture Media; Cytochrome P450; Deposition; Development; Drug toxicity; Endoderm; Enzymes; Evaluation; Exhibits; Extracellular Matrix; Fibroblast Growth Factor; Ficoll; Funding; Glycogen; Goals; Government; Hepatic; Hepatocyte; In Vitro; Indocyanine Green; Industrialization; Laminin; Legal patent; Lipids; Liver parenchyma; Low-Density Lipoproteins; Measures; Mesenchymal Stem Cells; Metabolism; Methodology; Methods; Monitor; Output; Persons; Phase; Pluripotent Stem Cells; Preclinical Drug Evaluation; Production; Property; Protocols documentation; Reagent; Reporter; Research; Resources; Screening procedure; Small Business Innovation Research Grant; Staging; Staining method; Stains; Stem cells; System; Technology; Testing; Therapeutic Studies; Toxicity Tests; Toxicology; Urea; Work; abstracting; activin A; base; cell type; drug discovery; improved; improved functioning; in vivo; induced pluripotent stem cell; macromolecule; meetings; new technology; novel; oncostatin M; phase 1 study; phase 2 study; sample fixation; scale up; tissue culture; two-dimensional; uptake

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.

描述（由申请人提供）： Primorigen Biosciences SBIR 提案摘要 Primorigen Biosciences 将利用 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来源的肝细胞的功能成熟度。该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，并鼓励细胞利用其自身的内源机制分泌和重塑细胞外基质，以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。第一阶段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分析来评估分化肝细胞的功能和成熟度。 II 期研究将验证和优化 II 期酶的化合物代谢和活性，并使改进的肝细胞分化方案适应更高通量的格式，例如 96 和 384 孔板，以便为药物发现和毒性研究进行化合物筛选。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商业化。 公共卫生相关性： Primorigen Biosciences, Inc. SBIR 项目叙述 全球范围内用于治疗和研究应用（包括药物发现和毒性筛选）的原代肝细胞短缺。此外，原代肝细胞在组织培养环境中倾向于丧失大部分肝功能，这表明非常需要可扩展且可重复的已知基因型肝细胞样细胞来源。虽然目前的干细胞培养方法已经成功且有效地产生了模仿原代肝细胞多种功能的肝细胞样细胞，但它们尚未成功地产生完全模仿原代细胞功能的细胞，特别是对毒性分析和关键肝功能至关重要的p450细胞色素酶的编码基因的表达。 SBIR技术将使用正在申请专利的专有培养基添加剂来解决这个问题，使干细胞能够利用其自身的内源性机制创造一个更像体内的微环境，从而产生功能更齐全的干细胞衍生肝细胞。