Thermoresponsive Poly(2-propyl-2-oxazoline) Surfaces of Glass for Nonenzymatic Cell Harvesting

As one of the nonenzymatic cell-harvesting technologies, a thermal-responsive surface based on poly(2-oxazoline)s has achieved initial success in supporting the adhesion and thermal-induced detachment of animal cells. However, because of the laborious preparation procedure, this technique was only limited to research purposes. In this work, through using poly(glycidyl methacrylate) (PGMA) as the anchor layer, poly(2-propyl-2-oxazoline)s (PPOx) were grafted onto glass wafers through a facile two-step coating and annealing procedure for nonenzymatic cell harvesting. In the first step, the piranha solution-activated glass wafers were immersed into the chloroform solution of PGMA and then annealed for a given period of time to immobilize PGMA onto the glass wafers through the bonding between epoxy groups and hydroxyl groups. In the second step, the PGMA-coated glass wafers were further immersed into the chloroform solution of carboxyl-functionalized PPOx. After annealing, PPOx were immobilized onto the PGMA layer through the bonding between carboxyl groups and the residual epoxy groups. Atomic force microscopy, X-ray photoelectron spectroscopy, and ellipsometry were used to characterize the modified glass wafers. The results of cytocompatibility evaluation showed that the PPOx-coated glass wafers were almost nontoxic and were able to support the adhesion and proliferation of L929 cells well. By lowering the temperature to 8 °C, L929 and Vero cells were successfully detached from the PPOx-coated glass wafers without any enzymatic treatment. Further cultivation has demonstrated that the cooling procedure had little effect on cell viability, and the cells still retained good viability after harvesting.