Primorigen Biosciences SBIR Proposal Abstract Primorigen Biosciences will develop a defined, xenobiotic-free synthetic cell culture matrix that fully supports maintenance and directed differentiation of human induced pluripotent stem cells (hiPSCs) down all three germ layers and dramatically reduces the costs of stem cell culture processes. The current, most commonly used matrices either create significant reliability and safety issues, or are effective only with certain cell lines given their limited mechanisms for cel adhesion. To address these drawbacks, Primorigen will develop a new synthetic cell culture matrix that recapitulates key attributes of natural extracellular matrix by presenting diverse synthetic ligands that offer numerous mechanisms for cell adhesion and facilitate sequestration of endogenous factors thereby reducing the need for and costs of using exogenous growth factors. Phase I studies will demonstrate feasibility by (a) creating a library of synthetic cell adhesion and endogenous factor sequestering ligands, (b) optimizing the coating conditions of ligand mixtures onto polystyrene surfaces, and (c) validating the new synthetic cell culture matrix for hiPSC maintenance and directed differentiation into various lineages. Phase II objectives will build on this work by expanding the synthetic ligand mixture approach to validate their use in large scale production and differentiation of hiPSCs. Commercial outputs will include (i) a broadly applicable synthetic Matrigel replacement comprised of synthetic modular ligand mixtures optimized for stem cell maintenance and differentiation, (ii) polystyrene tissue culture platforms (e.g., Petri dish, multiwell plate and microcarrier) pre-coated with the synthetic modular ligand mixtures, and (iii) a library of synthetic modular ligands from which made to order, customized mixtures can be assembled according to individual customers' unique needs.
Public Health Relevance Statement: Public Health Relevance: Primorigen Biosciences SBIR Proposal Narrative The self-renewal and differentiation capacity of genetically stable human pluripotent stem cells create enormous potential for advancing drug screening and creating new cell therapies for a wide variety of tragic degenerative diseases. Current protocols for maintenance and differentiation of pluripotent cells use undefined, cell culture matrices of animal origin, which present lot-to-lot variability and safety issues. Defined, xenobiotic-free matrices have been developed that address these problems, but their use is limited to fewer cell lines than observed with undefined matrices. To address these combined shortcomings, Primorigen will develop a synthetic, defined, xenobiotic- free cell culture matrix from peptide mimics of several key ECM components and growth factors to provide multiple adhesion ligands to the cells and facilitate binding and temporal release of growth factors. The resulting product will ensure broad applications across a wider diversity of cell lines and facilitate differentiation down all three grm layers, while reducing the need for exogenous growth factors to drive down costs of stem cell culture processes. The xenobiotic-free attributes of the matrix will facilitate cell therapy applications, and the substantial costs savings resulting from lower exogenous growth factor use will be compounded in cGMP production, greatly increasing the affordability of new stem cell therapies.
Project Terms: abstracting; Address; Adherent Culture; Adhesions; AFP gene; Albumins; Amino Acid Sequence; Animals; Binding (Molecular Function); Biological Sciences; Cardiac Myocytes; Cell Adhesion; Cell Culture Techniques; Cell Differentiation process; cell growth; Cell Line; Cell Lineage; Cell Maintenance; Cell Therapy; Cell-Cell Adhesion; Cells; cGMP production; Clinical; Complex Mixtures; cost; Cost Savings; culture plates; Cyclic GMP; Degenerative Disorder; design; Differentiation Antigens; Disadvantaged; Ectoderm; Endoderm; Endogenous Factors; Ensure; Evaluation; Extracellular Matrix; Extracellular Matrix Proteins; Funding; Germ Layers; Goals; Government; Growth Factor; Heparin Binding; Hepatocyte; Human; Individual; induced pluripotent stem cell; interest; Karyotype; large scale production; Libraries; Ligand Binding; Ligands; Maintenance; matrigel; Mesoderm; Morphology; Mus; nerve stem cell; nestin protein; novel; Output; pathogen; Peptides; Performance; Persons; Phase; phase 1 study; phase 2 study; pluripotency; Pluripotent Stem Cells; Polystyrenes; Preclinical Drug Evaluation; Process; Protocols documentation; public health relevance; Regenerative Medicine; Research; Research Personnel; Safety; sarcoma; self-renewal; Small Business Innovation Research Grant; stage-specific embryonic antigen 4; stem cell therapy; Stem cells; Surface; surface coating; Technology; Testing; tissue culture; Translations; Work; Xenobiotics
