Abstract

Defined xeno-free laminin matrix-based differentiation protocols for making clinical quality stem cell -derived cells for regenerative medicine

Karl Tryggvason, Professor, Duke-NUS Graduate Medical School & Karolinska Institutet, Stockholm

The mainstream of human stem cell research is currently moving toward the development of methods for differentiation of stem cells to different cell types that can be used in regenerative medicine. Stem cell -derived cells hold great promise for the treatment of various diseases, such as heart disease, type I diabetes, Parkinson’s disease, muscular dystrophy, vascular and liver disease and macular degeneration, as well as connective tissue diseases. A prerequisite for high clinical quality cells is that they have been established, differentiated, expanded and maintained under chemically defined conditions and that the cell culture media are devoid of animal derived products. The methods should fulfill GMP conditions. Several differentiation protocols have been reported, but most of them do not fulfill those criteria. During development in vivo, expression of appropriate transcription factors, presence of suitable growth factors and hormones, as well as biologically correct cell-cell and cell-matrix interactions, are crucial for the differentiation process. All highly organized cell types in the body such as epithelial and endothelial cells, muscle fibers, nerves, and adipocytes are associated with an ultrathin basement membrane (basal lamina) which contains special collagen molecules and proteoglycans, as well as over 15 different laminin isoforms that are important for cell adhesion, differentiation, migration and resistance to apoptosis. Most of the laminins are cell type specific, suggesting their importance for development and maintenance of specific cell types. Our group has cloned and expressed most human laminins and studied them for effects in differentiation and phenotype stability. Here, results describing robust methods for development and characterization of cardiomyocytes and endothelial cells will be described.