Preliminary report on cell culture on a thermally reversible copolymer

Anchorage-dependent cells (NRK 49F) were grown at 37 degrees C in serum containing medium on glass coated with a N-isopropyl acrylamide/N-t-butyl acrylamide copolymer (NIPAaM/NTBAM) with a lower critical solution temperature (LCST) of 8 degrees C. After incubating the dishes and adherent cells at 4 degrees C for 1 h, the cells had started to detach in 'sheets', without the use of trypsin; by 2 h the cells had fully detached and formed aggregates. We presume that at 4 degrees C, below the LCST of the coating, the polymer swelled and eventually dissolved, leaving the cells unattached to the solid substrate. We speculate that the 'sheet' morphology of the detaching cells implies that the extracellular matrix produced by the cells during culture was left intact, which may then alter the nature of their subsequent behaviour. The polymer was not optimum for cell attachment and optimization would be needed to realize the potential of this novel mode of cell culture and recovery.