Retinal pigmented epithelium cultures on thermally responsive polymer porous substrates.

ABSTRACT

A cross-linkable co-polymer of UV-sensitive 4-(N-cinnamoylcarbamide)methylstyrene (CCMS) and N-isopropylacrylamide (NIPAAm), was applied to porous tissue culture inserts. Surface chemical analyses of the inserts show an introduction of a thermally responsive polymer comparable to that on similarly incorporated non-porous polystyrene surfaces. Contact angle measurements as well as atomic force microscopy show a surface change in response to changing temperature in an aqueous environment, from hydrophilic, extended polymer chains below 32 degrees C to a dense hydrophobic film above 32 degrees C. Cell growth on porous inserts allowed measurement of cell expression, such as transepithelial resistance and fluid transport, which are not observable on cells from non-porous surfaces. Cultures of retinal pigmented epithelium (RPE) were able to restore an environment similar to in vivo by forming a tight junction barrier membrane upon confluence at 37 degrees C, as observed by changes in morphology, transepithelial resistance, and directionally-specific fluid transport. In addition, cells cultured on these surfaces detached as an oriented polarized sheet when the inserts were brought to 20 degrees C. This cell sheet was transplanted to other tissue culture surface without polymer detachment or dissolution, or cell damage caused by traditional detachment methods using proteolytic enzymes.