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Successive Grafting of PHEMA and PIPAAm onto Cell Culture Surface Enables Rapid Cell Sheet Recovery
김선종,김원일,Masayuki Yamato,Teruo Okano,Akihiko Kikuchi,권오형 한국조직공학과 재생의학회 2013 조직공학과 재생의학 Vol.10 No.3
Fabrication of functional tissue constructs from designed three-dimensional structure of cells using layered method of cultured cell sheets could prove to be an attractive approach to tissue engineering. Rapid recovery of cell sheets is considered important to maintain the biological functions and viability of recovered cell sheets, as well as for practical assembly of tissue structures. To accelerate required culture substrate hydrophilic/hydrophobic structural changes in response to culture temperature alteration, different amounts of poly(2-hydroxyethyl methacrylate) (PHEMA) and poly(N-isopropylacrylamide) (PIPAAm) were grafted successively onto tissue culture polystyrene (TCPS) dishes by electron beam irradiation. Analysis by attenuated total reflection-Fourier transform infrared revealed that PHEMA and PIPAAm were successfully grafted to the surfaces of TCPS dishes. PIPAAm-PHEMA-grafted TCPS (PIPAAm-PHEMA-TCPS) dishes were compared with PIPAAm-grafted TCPS (PIPAAm-TCPS) dishes for cell sheet detachment experiments. Approximately 75 min was required to completely detach cell sheets from PIPAAm-TCPS dishes, compared to only 13 min to detach cell sheets from PIPAAm-PHEMA(40 wt%)-TCPS dishes, which is successively grafted with 40 wt% of HEMA and 60 wt% IPAAm solutions. PHEMA is a well-known as a highly hygroscopic polymer. In the case of PIPAAm-PHEMA-TCPS dish, PHEMA layer acted as a water pool to accelerate the hydration of PIPAAm layer due to the effective and simultaneous water supply to PIPAAm layer, resulting in rapid hydration of grafted PIPAAm molecules and detachment of cell sheet compare to PIPAAm-TCPS dishes.