METHOD FOR FORMATION OF 3 DIMENTIONAL MULTICELLULAR SPHEROIDS USING RGD-FUNCTIONALIZED ELASTIN-LIKE MULTIBLOCK BIOPOLYMERS AND FORMATION MANUFACTURING THE SAME

摘要

PURPOSE: A method for forming three-dimensional cell aggregate using an elatine-like artificial extracellular substrate, and the three-dimensional cell aggregate are provided to safely enhance productivity without cytotoxicity, and to ensure tissue regeneration ability and biocompatibility. CONSTITUTION: A method for forming three-dimensional cell aggregate containing an elastine-like substrate protein comprises: a step of chemically synthesizing 4 oligonucleic acid salt for forming 7 pentapeptide VGRGD(VGVPG)_6 unit genes; a step of replicating ELP[RGD-V_6]_n(100); a step of expressing E. coli BLR(DE3) in 40 liter TB(terrific broth); and a step of performing reverse phase transition for the purification in a PBS solution at pH 7.2. The three-dimensional aggregate containing an elastine-like artificial extracellular substrate protein is made of a multiple RGD material for integrin binding on the surface of a cell. The multiple RGD material is ELP[RGD-V_6]_n. [Reference numerals] (100) N2a cell culture; (201) Protein matrix specimen and protein density measurement; (203) Characteristic analysis; (205) Surface shape and roughness measurement; (207) Cell adhesion and affinity measurement; (209) Cell diffusion and scratch movement analysis; (211) Neuronal differentiation measurement of N2a cells; (213) Cell survivality measurement