| Project/Area Number |
04453108
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
高分子物性・高分子材料（含機械材料）
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| Research Institution | Tokyo Women's Medical College |
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Principal Investigator |
OKANO Teruo Tokyo Women's Medical College, Medical, Professor, 医学部, 教授 (00130237)
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| Co-Investigator(Kenkyū-buntansha) |
KIKUCHI Akihiko Tokyo Women's Medical College, Medical, Assistan, 医学部, 助手 (40266820)
SUZUKI Ken Tokyo Women's Medical College, Medical, Assistan, 医学部, 助手 (90216375)
山田 則子 東京女子医科大学, 医学部, 助手 (50107314)
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| Project Period (FY) |
1992 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥7,600,000 (Direct Cost: ¥7,600,000)
Fiscal Year 1994: ¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1993: ¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1992: ¥3,400,000 (Direct Cost: ¥3,400,000)
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| Keywords | Poly (isopropylacrylamide) / Polymer grafted surface / Temperature responsive surface / Control of cell detachment / Mechanism of cell detachment / Intelligent materials / 界面応答性材料 / ポリイソプロピルアクリルアミド(PIPAAm) / 表面グラフト / 肝細胞 / 内皮細胞 / 細胞脱着 / 細胞回収 / 親水-疎水性の表面変化 |
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| Research Abstract |
Abstract :
Poly (N-isopropylacrylamide) (PIPAAm) shows reversible hydrophilic / hydrophobic property changes upon temperature changes. We have utilized this polymer for hydrophilic / hydrophobic surface property alteration for novel thermo-responsive recovery system for cultured cells. Cultured cells can easily be recovered from the surfaces of PIPAAm-grafted culture dishes by simple temperature decreases. In this report detachment mechanism of cultured cells by lowering temperature treatment is discussed. PIPAAm molecules are introduced to polystirene culture dishes by electron-beam irradiation. Used cells are bovine aortic endothelial cells and rat hepatocytes and cultured in a humidified atmosphere of 5% CO_2 at 37ﾟC.
The effect of temperature on recovery of cultured cells from PIPAAm-grafted dish was investigated with respect to both degree of surface hydrophilic / hydrophobic change and cell metabolism change. Greater recovery of cultured cells can be achieved by incubation at 25ﾟC for 5 min after 30 min incubation at a definite temperature (4-25ﾟC). Recovery of cells by temperature treatment was decreased by treating cultured cells with sodium azide, implying that cellular metabolism is involved in the detachment process of cultured cells. Recovered cells by lower temperature treatment well proliferated on tissue culture dishes and retained high functionality as judged by secretion of albumin from hepatocytes compared with cells recovered by conventional trypsin treatment. Confluent culture of cells could also be recovered by temperature treatment. This technique might be useful for construction of artificial tissues such as artificial skin.
These results strongly suggest the feasibility of novel cell recovery system using temperature-responsive hydrophilic / hydrophobic alterations of PIPAAm-grafted surfaces.
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