SBIR-STTR Award

Primorigen Biosciences LLC SBIR Project Abstract Primorigen will use SBIR funds to develop and validate a microcarrier-based cell culture system for selection, scale-up and feeder-free mass production of iPS cells, based on Primorigen's proprietary pluripotent stem cell attachment factor (StemCadhere") for direct physical selection and propagation of iPSC. Phase I studies will define coupling chemistry that optimizes E-cadherin mediated cell adhesion to StemCadhere"-coated microcarriers, analyze iPSC attachment efficiency and propagation on the StemCadhere"-coated microcarriers, analyze the utility of StemCadhere-coated microcarriers for direct physical selection of newly induced iPS cells, and compare performance of cryopreserved iPSC on StemCadhere"-coated microcarriers with Matrigel-coated microcarriers. This work will provide the foundation for Phase II studies, which will investigate production of differentiated cells directly on the microcarriers. Similar methods will be used to test and select cellular adhesion molecules specific for each germ layer (ectoderm, mesoderm and endoderm), and for a limited number of terminally differentiated lineages such as hepatocytes, cardiomyocytes, and hematopoietic cells. The initial commercialized products will be a) ready-to-use (cryopreserved) iPS cells attached to the microcarriers, ready for addition of differentiation factors and downstream protocols;and b) a new application for attachment factors coupled to microcarriers that Primorigen can market at large to pharmaceutical and biotechnology firms engaged in differentiated cell production from iPS.

Public Health Relevance:
Primorigen Biosciences LLC SBIR Project Narrative Current methods for production of iPS cells require labor intensive manual culturing steps, frequent feeding of cultures, and inclusion of drug-selectable resistance markers to achieve purification of iPS from non-induced parental cells (such as foreskin fibroblasts). To fulfill the promise of iPS cells as a source for cellular therapies and differentiated cells for pharmaceutical development, it is mandatory to avoid the use of lentiviral vectors and other agents capable of genomic integration, which precludes the use of integrated drug-resistance markers for iPS selection. In addition, to decrease variability in production lots, it is desirable to automate as much of the culture as possible, while maintaining the proliferation and pluripotency of the iPS cells. Primorigen's SBIR proposal will address this need by developing a comprehensive, integrated solution for selection and mass production of iPS cells by taking advantage of Primorigen's proprietary stem cell attachment factor for feeder-free propagation of iPS cells in microcarrier suspension culture.

Thesaurus Terms:
Address;Adhesion Molecule;Adhesions;Assay;Bioassay;Biologic Assays;Biological Assay;Bioreactors;Biotechnology;Blood Coagulation Factor Iv;Blood Serum;Cam 120/80;Ca++ Element;Cadherin-1;Calcium;Cardiac Myocytes;Cardiocyte;Cell Adhesion;Cell Adhesion Molecules;Cell Attachment;Cell Culture System;Cell Line;Cell Lines, Strains;Cell-Extracellular Matrix;Cell-Matrix Adhesions;Cell-Matrix Junction;Cellline;Cells;Cellular Adhesion;Chemistry;Clinical;Coagulation Factor Iv;Conditioned Culture Media;Conditioned Medium;Coupled;Coupling;Cryofixation;Cryopreservation;Culture Media, Conditioned;Development;Dysembryoma;E-Cadherin;Ecm;Ectoderm;Endoderm;Epithelial Calcium-Dependent Adhesion Protein;Epithelial-Cadherin;Evaluation;Extracellular Matrix;Factor Iv;Foundations;Funding;Generations;Germ Layers;Government;Harvest;Heart Myocyte;Hematopoietic;Hepatic Cells;Hepatic Parenchymal Cell;Hepatocyte;Human;Human, General;Immunoassay;Karyotype;Letters;Liver Cells;Maintenance;Maintenances;Mammals, Mice;Man (Taxonomy);Man, Modern;Marketing;Mediating;Mesoderm;Methods;Mice;Monitor;Mother Cells;Murine;Mus;Muscle Cells, Cardiac;Muscle Cells, Heart;Myocytes, Cardiac;Performance;Persons;Pharmaceutical Agent;Pharmaceuticals;Pharmacologic Substance;Pharmacological Substance;Phase;Pluripotent Stem Cells;Polystyrenes;Polystyrol;Process;Production;Progenitor Cells;Proteins;Protocol;Protocols Documentation;Publications;Recombinants;Recovery;Regenerative Medicine;Reporting;Research;Sbir;Sbirs (R43/44);Science Of Chemistry;Scientific Publication;Serum;Small Business Innovation Research;Small Business Innovation Research Grant;Somatic Cell;Source;Staging;Stem Cells;Suspension Culture;Suspension Substance;Suspensions;System;System, Loinc Axis 4;Teratoid Tumor;Teratoma;Testing;Tumor-Derived;Uvomorulin;Wisconsin;Work;Xenobiotics;Abstracting;Base;Cardiac Myocyte;Cardiomyocyte;Cell Adhesion Protein;Cold Preservation;Cold Storage;Commercialization;Cultured Cell Line;Gene Product;Hesc;Human Es Cell;Human Esc;Human Embryonic Stem Cell;Induced Pluripotent Stem Cell;Karyogram;Matrigel;Medical Schools;Novel;Phase 1 Study;Phase 2 Study;Pluripotency;Regenerative;Scale Up;Stem Cell Population;Suspension

Primorigen Biosciences (R) Abstract Primorigen Biosciences will use Phase II SBIR funds to advance its novel, 3-dimensional system for robust expansion of induced pluripotent stem cells (iPSCs) by a) scaling up iPSC production in 3L bioreactors, b) validating the bioreactor-expanded iPSCs for differentiatation into lineages of value for pharmaceutical drug screening, and c) validating large-scale cryopreservation of the iPSCs. Phase I studies successfully demonstrated feasibility of iPSC expansion in the new system, with the added benefits of substantially lowering media and labor requirements versus standard 2-dimensional culture. Phase II studies will validate the system for larger scale applications as described above. The ultimate commercial goals are to enable industrialization of stem cells and their derivatives by addressing the current technical and market need for cost-effective large scale production of pluripotent stem cells, a key barrier to the use stem cells for widespread drug screening and clinical therapeutics. To ensure utility for both applications, Phase II studies will continue the work initiated in Phase I to make the system xeno-free and adapatable for ultimate GMP production. The initial production applications will be targeted to companies requiring large quantities of phenotypically relevant cell types for drug discovery. Longer term, the system's paradigm-changing efficiency and scalability will be adapted to produce clinical-grade therapeutic cells derived from stem-cells.

Public Health Relevance Statement:


Public Health Relevance:
Primorigen Biosciences(R) Project Narrative To fulfill the promise of iPS cells as a source for cellular therapies and differentiated cells for pharmaceutical drug evelopment, the market demands cost-effective and scalable methods for large-scale iPSC production. To minimize batch-to-batch variability, the culture process should be automated to the maximum extent while maintaining the proliferation and pluripotency of the iPS cells. Primorigen's SBIR proposal will address these needs by developing a comprehensive, integrated solution for feeder-free, xeno-free, three-dimensional mass production of iPS cells.

Project Terms:
3-Dimensional; abstracting; Address; Adhesions; base; Biological Products; Bioreactors; Cardiac Myocytes; Cell Culture Techniques; Cell Proliferation; cell type; Cells; Clinical; clinical application; Communication; Consumption; cost; cost effective; Cryopreservation; density; Development; drug discovery; Drug Formulations; Endoderm; Ensure; Evaluation; Freeze Drying; Funding; Future; Goals; Government; Hematopoietic; Hepatocyte; induced pluripotent stem cell; Industrialization; Industry; Investigation; Laboratory culture; large scale production; Marketing; Methods; neurodevelopment; novel; Nutrient; Oxygen; Persons; Pharmaceutical Preparations; Pharmacologic Substance; Phase; phase 1 study; phase 2 study; pluripotency; Pluripotent Stem Cells; Preclinical Drug Evaluation; Process; Production; Proteins; public health relevance; Recovery; relating to nervous system; Research; Running; scale up; screening; Shipping; Ships; Small Business Innovation Research Grant; Solutions; Source; stem; Stem cells; Supplementation; Suspension Culture; Suspension substance; Suspensions; System; Technology; Testing; Therapeutic; Toxic effect; two-dimensional; Vitronectin; Wood material; Work; Xeno; Xenobiotics