Uses of thermoresponsive and RGD/insulin-modified poly(vinyl ether)-based hydrogels in cell cultures

Abstract

Thermoresponsive hydrogels were synthesized by radiation copolymerization of ethylene glycol vinyl ether (1) and butyl vinyl ether (2) in the presence of cross-linking agent diethylene glycol divinyl ether. The comonomer ratio (monomer 1/monomer 2) and the cross-linker concentration were kept constant at 60 :40 (mole percentage in the monomeric mixture) and 4% (mole basis), respectively. The hydrogels showed a volume-phase transition in the temperature range 10-25 ° C and their swelling behaviour was reversible. The gels were modified by a cell adhesion factor, the RGD sequence of fibronectin, and a cell growth factor, insulin. However, they lost their thermoresponsive character after modification. The use of the gels in cell culture was investigated without using a proteolytic enzyme or serum. Cell culture studies realized by human skin fibroblasts (HS An1) showed that the cells can attach and proliferate on the surface of a thermoresponsive polymer. 80% of the cultured cells were readily detached from the polymer surface by lowering the incubation temperature from 37 ° C to 10 ° C for 30 min. In the studies carried out with RGD or insulin-modified hydrogels in serum-free cultures, higher values of cell proliferation (9 × 105 cells/ml) were obtained on the insulin-modified hydrogels, whereas higher values of cell attachment were obtained on the RGD-immobilized surfaces.