Efficacy of human stem/primary cells for biomedical applications hinges upon successful in vitro culture and manipulation. Cell attachment and growth represent critical steps that dictate the overall culture success. To promote cell attachment and growth, animal or human-origin extracellular matrix (ECM) proteins are widely used. ECM not only provides structural support to the cells in culture, but also presents intricate biologically active motifs to dictate cell function. However, whole ECM protein preparations run the risk of pathogen or immunogen transmission to the cells, and are associated with considerable lot-to-lot variability. There is a compelling need for a chemically defined synthetic platform for robust human cell culture. Many companies are developing xeno-free fully chemically defined culturing media. Since there are no fetal bovine serum (FBS) and other ECM proteins in these new culture media, xeno-free coatings on culturewares surfaces will be critically important for human cells to attach and grow. To answer this call, StemCellLife LLC aims to develop low cost and xeno-free, highly active pure synthetic coatings that are applicable to regular cell culturewares based upon immobilization of ECM-derived short adhesive peptides in looped structures to mimic the physical and biochemical properties of the natural ECM. Instead of whole ECM proteins, short adhesive peptides that act as functional cell binding domains in the whole ECM molecules will be used in the coatings. Although several short peptide sequences have been identified, few of them from the existing commercial sources can support the adhesion, growth, and long-term culture of human stem cells, such as human neural stem/progenitor cells (hNSCs), human embryonic stem cells (hESCs), and human induced pluripotent stem cells (hiPSCs), and human primary cells. The overall goal of this SBIR Phase I project is to establish surface treatment technologies that allow stable immobilization and full exposure of functional cell binding domains of short peptides on regular cell cultureware surfaces to support human cell culture and function. Our central hypothesis is that a novel technology, which allows stable immobilization and full exposure of functional cell binding domains of short adhesive peptides on cultureware surfaces, would support the survival, adhesion, growth, and long-term culture of human cells. In this Phase I project, a series of short peptide-based 100% synthetic coating products will be developed for the culture of human neural stem/progenitor cells (hNSCs), human primary neurons, human primary endothelial cells, and human primary epithelial cells. In the future Phase II project, we will develop 100% synthetic coating products to support the long-term culture of hiPSCs, hESCs, and other cells. Our proposed project to create chemically defined synthetic platform for robust human cell culture based upon xeno-free and highly active pure synthetic coatings of ECM-derived short adhesive peptides will fill in the gap in current product development to effectively support human cell survival and function. The platform will advance all areas of human cell research and therapy development, and direct future efforts for human tissue regeneration and drug discovery. By eliminating the need for whole ECM molecules, the highly active pure synthetic coatings under development in this project will greatly simplify human cell culture with enhanced efficacy and lower cost. Development of the products will generate major economic and social impacts.
This project aims to create chemically defined synthetic platform for robust human stem/primary cell culture based upon xeno-free and 'highly active pure synthetic coatings' of ECM-derived short adhesive peptides in looped structures that are applicable on regular cell cultureware surfaces. The platform will advance all areas of human cell research and therapy development, and direct future efforts for human tissue regeneration and drug discovery. By eliminating the need for whole ECM molecules from animal or human sources, the 'highly active pure synthetic coating' products under development in this project will greatly simplify human cell culture with enhanced efficacy and lower cost. Development of the products will generate major economic and social impacts.
