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.
Primorigen Biosciences(R) Project Narrative Current culture and differentiation of hPSCs depends upon the use of supraphysiological concentrations of a variety of growth factors, a process that results in inordinately high costs for expansion and differentiation and does not replicate signaling in the in vivo microenvironment. Culture systems that can amplify the activity of exogenous factors and harness the activity of endogenous growth factors are needed to provide better in vitro models for study of physiological growth factor roles and more economical methods for producing differentiated cell types from expanded pluripotent cell stocks. The economics of production is particularly important for disease modeling, drug screening, and clinical applications. To address this need, Primorigen will develop a paradigm-changing technology based on a novel heparin-binding peptide (HEPpep) that dramatically reduces costs by enhancing the impact of endogenous growth factors and minimizing the need for exogenous growth factors for expansion and directed differentiation of human pluripotent stem cells (hPSCs). This project will demonstrate the use of HEPpep on standard polystyrene culture vessels, for expansion of hPSCs under reduced growth factor conditions, and by using Primorigen's proprietary, high efficiency hepatocyte differentiation protocol for differentiation o hiPSCs on HEPpep modified surfaces, providing a proof of principle for extending the technology to other lineages.