Cell detachment and preservation of hematopoietic stem cells on thermo-responsible pluronicimmobilized membranes

Long-term (7 days) cell culture was successively performed on the Pluronic-immobilized flask in the current investigation. The Pluronic F127 and F68, a triblock copolymer of ethylene oxide, and propylene oxide was activated using carbonyldiimidazole (CDI), and CDI-activated Pluronic was subsequently immobilized on the surface of a lysine-coated polystyrene tissue culture flask. L929 cells were successively detached from the Pluronic-immobilized flask by cooling the flask to 4-15°C. This detachment is due to the hydration and dehydration properties of Pluronic F127, depending on the temperature. Furthermore, we show that hematopoietic stem cells in umbilical cord blood are extensively preserved in the Pluronic-immobilized flask under bio-inert conditions compared to those in commercially available tissue culture flask. We anticipate that this is caused by the flexible brush-like segments of Pluronic on the tissue culture flask. The flexible segments of Pluronic induce hematopoietic stem cells to bio-inert cultivation of hematopoietic stem cells.