CELL ADHESION PEPTIDES TO IMPROVE CULTURE SURFACE

NUMEROUS ANCHORAGE-DEPENDENT CELLS SHOW IMPROVED ATTACHMENT,IMPROVED GROWTH, AND REDUCED SERUM REQUIREMENTS WHEN CULTURED ON SURFACES COATED WITH EXTRACELLULAR MATRIX COMPONENTS (ECM'S). THE MAJOR ACTIVE PROTEINS IN ECM'S ARE FIBRONECTIN, LAMININ, AND TYPE IV COLLAGEN. MOST OF THE CELL ADHESION ACTIVITIES OF THESE PROTEINS RESIDE IN TWO PEPTIDE SEGMENTS (RGD AND YIGSR), AND BOTH PEPTIDES ARE ACTIVE WHEN IMMOBILIZED ONTO SURFACES. IT IS THEREFORE REASONABLE THAT THESE TWO PEPTIDES MIGHT BE IMMOBILIZED TO PRODUCE CELL CULTURE SURFACES WITH PROPERTIESSIMILAR TO THOSE OBSERVED WITH ECM'S. THIS RESEARCH PROPOSES TO COVALENTLY IMMOBILIZE THESE PEPTIDES ONTO POLYSTYRENE, WITH THE EXPECTATION OF PRODUCING INEXPENSIVE TISSUE CULTURE PRODUCTS WITH GREATLY IMPROVED CELL CULTURE PROPERTIES. IN PHASE I, THESE PEPTIDES WILL BE IMMOBILIZED USING (3)H-LABELS TO QUANTITATE IMMOBILIZATION. THEN, SIX REPRESENTATIVE CELL TYPES WILL BE USED TO ASSAY FOR CELL ATTACHMENT AND CELL GROWTH IN BOTH LOW AND NORMAL SERUM LEVELS. IN PHASE II, SEVERAL PARAMETERS WILL BE EVALUATED: (1) ATTACHMENT AND GROWTH IN SERUM-FREE MEDIA, (2) MAINTENANCE OF DIFFERENTIATED CELL CHARACTERISTICS, AND (3) GENERATION OF SECONDARY CELL PRODUCTS. PROOF OF CONCEPT OF THIS TECHNOLOGY SHOULD GREATLY IMPROVE THE EFFICIENCY OF CULTURING MANY TYPES OF ANCHORAGE-DEPENDENT ANIMAL CELLS.