Growth factor release from thermally reversible tissue culture substrates.

Thermally reversible poly(N-isopropylacrylamide) (PIPAAm) was covalently grafted onto tissue culture dishes to allow detachment of cultured cells upon temperature change from physiological to room temperature. In addition the grafted polymer matrix was used to entrap biomolecules such as growth factors either to be released by diffusion early in cell cultures, or remain entrapped and be reversibly exposed to cell receptors. Experiments with model proteins trypsin and insulin show that amount loaded and released depends upon the PIPAAm grafting density. Dishes grafted with 2.5 microgram/cm2 PIPAAm released approximately four times more model protein over 4 h than dishes grafted with 1.8 microgram/cm2. This in vitro drug delivery system can be used to deliver factors to the basal side of cells early in cell culture by providing high local concentrations without high bulk concentration. Cultures of human retinal pigmented epithelium showed higher growth rate on insulin loaded dishes than on controls containing a similar bulk solution concentration. These cultures retained the ability to detach singly or as confluent sheets from the loaded surfaces.