Design of antioxidative biointerface for separation of hematopoietic stem cells with high maintenance of undifferentiated phenotype

During cell cultivation, excessively generated reactive oxygen species (ROS) affect cellular properties and functions. Although cell cultivation media contain several types of low-molecular-weight antioxidants, these small antioxidants are internalized into the mitochondria and they disrupt regulated redox balance. Here, we developed a novel biointerface that effectively eliminates ROS on a cell culture surface. Poly(ethylene glycol)-b-poly[4-(2,2,6,6-tetramethylpiperidine-1-oxyl)aminomethylstyrene] (PEG-b-PMNT) was synthesized and covalently coated on a carboxyl group-activated culture dish using sec-amino groups on a PMNT segment followed by immobilization of anti-CD34 antibodies. CD34-positive hematopoietic stem progenitor cells (HSPCs) were separated from mice fetal liver cells using our polymer-coated cell culture dish. The separated HSPCs possessed intact mitochondrial membrane potential compared with those in the conventional cell cultivation system. In addition, the expression level of CD34 was maintained for an extended period on our culture dish with the antioxidative biointerface. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 2080-2085, 2016.