A Novel Substrate to Amplify Growth Factor Impact on hPSC Differentiation

描述（由申请人提供）：Primorigen Biosciences（R）项目摘要Primorigen将使用SBIR基金开发一种范式改变技术，用于使用新型肽扩增和定向分化人类多能干细胞（hPSC），该肽通过放大内源性生长因子的影响并最大限度地减少对外源性生长因子的需求来显著降低成本。该肽已被用于放大多种生长因子的生理效应，通过提案中描述的机制。I期研究将证明肽在标准聚苯乙烯培养容器上的效用，以增强hiPSC在含生长因子减少的无血清培养基中的增殖，并作为更经济有效的定向分化的有效试剂。我们将 通过使用Primorigen专有的高效肝细胞分化方案在肽修饰的表面上分化hiPSC，证明了后者的可行性，为将该技术扩展到其他谱系提供了原理证明。II期活动将测试和验证肽用于1）在搅拌细胞悬浮生物反应器中的微载体上按比例放大生产hiPSC，2）在肽存在下其他谱系的有效生长因子依赖性分化，以伴随按比例放大。根据该提案开发的产品将包括肽包被培养皿沿着，以及用于hiPSC维持和分化为各种细胞类型（从肝细胞开始）的减少生长因子培养基。

DESCRIPTION (provided by applicant): Primorigen Biosciences(R) Project Abstract Primorigen will use SBIR funds to develop a paradigm-changing technology for expansion and directed differentiation of human pluripotent stem cells (hPSCs) using a novel peptide that dramatically reduces costs by amplifying the impact of endogenous growth factors and minimizing the need for exogenous growth factors. The peptide has been used to amplify the physiological effects of multiple growth factors, through mechanisms described in the proposal. Phase I studies will demonstrate the utility of the peptide on standard polystyrene culture vessels to enhance proliferation of hiPSCs in reduced growth factor-containing serum-free media, and to serve as an effective agent for more cost-efficient directed differentiation. We will demonstrate the latter feasibility by using Primorigen's proprietary, high efficiency hepatocyte differentiation protocol for differentiation of hiPSCs on the peptide-modified surfaces, providing proof of principle for extending the technology to other lineages. Phase II activities will test an validate the peptide for use in 1) scaled-up production of hiPSCs on microcarriers in stirred-cell suspension bioreactors, 2) effective growth-factor dependent differentiation of other lineages in the presence of the peptide, to accompany scale-up. Products developed from this proposal will include the peptide-coated cultureware along with companion, reduced growth factor media for hiPSC maintenance and differentiation into various cell types, beginning with hepatocytes.