Rapid harvesting of stem cell sheets by thermoresponsive bulk poly(N-isopropylacrylamide) (PNIPAAm) nanotopography

Abstract

To date, cell sheet engineering-based technologies have actualized diverse scaffold-free bio-products to revitalize unintentionally damaged tissues/organs, including cardiomyopathy, corneal defects, and periodontal damage. Although substantial interest is now centered on the practical utilization of these bio-products for patients, the long harvest period of stem cells- or other primary cell-sheets has become a huge hurdle. Here, we dramatically reduce the total harvest period of a cell sheet (from cell layer formation to cell sheet detachment) composed of human bone marrow mesenchymal stem cells (hBMSCs) down to 2 d with the help of bulk thermoresponsive poly(N-isopropylacrylamide) (PNIPAAm) substrate nanotopography, which is not achievable via the previous grafting methods using PNIPAAm. We directly replicated an isotropic 400 nm-nanopore-array pattern on a bulk PNIPAAm substrate through UV polymerization of highly concentrated NIPAAm monomers, which was achieved using a remarkably increased Young's modulus of bulk PNIPAAm that was 1500 times higher than conventional PNIPAAm. The rapid harvesting of the hBMSC sheet on the bulk PNIPAAm substrate nanotopography was not only based on the accelerated formation and maturation of the hBMSC layer, but also the easy detachment of the hBMSC sheet induced by the abrupt change in the surface roughness of the substrate below the lower critical solution temperature (LCST) owing to the enlarged surface area of the substrate. Our findings may contribute to reverse presumptions about the limitations regarding the grafting methods for the cell sheet harvest and could broaden the practical utilization of cell sheets for patients in the near future.