Temperature-Responsive Cell Culture Surfaces Enable "On-Off" Affinity Control between Cell Integrins and RGDS Ligands
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- 作者：Mitsuhiro Ebara ; Masayuki Yamato ; Takao Aoyagi ; Akihiko Kikuchi ; Kiyotaka Sakai ; and Teruo Okano
- 刊名：Biomacromolecules
- 出版年：2004
- 出版时间：March 2004
- 年：2004
- 卷：5
- 期：2
- 页码：505 - 510
- 全文大小：111K
- 年卷期：v.5,no.2(March 2004)
- ISSN：1526-4602
文摘
In this study, specific interactions between immobilized RGDS (Arg-Gly-Asp-Ser) cell adhesion peptidesand cell integrin receptors located on cell membranes are controlled in vitro using stimuli-responsive polymersurface chemistry. Temperature-responsive poly(N-isopropylacrylamide-co-2-carboxyisopropylacrylamide)(P(IPAAm-co-CIPAAm)) copolymer grafted onto tissue culture grade polystyrene (TCPS) dishes permitsRGDS immobilization. These surfaces facilitate the spreading of human umbilical vein endothelial cells(HUVECs) without serum depending on RGDS surface content at 37 
C (above the lower critical solutiontemperature, LCST, of the copolymer). Moreover, cells spread on RGDS-immobilized surfaces at 37 Cdetach spontaneously by lowering culture temperature below the LCST as hydrated grafted copolymer chainsdissociate immobilized RGDS from cell integrins. These cell lifting behaviors upon hydration are similar toresults using soluble RGDS in culture as a competitive substitution for immobilized ligands. Binding of cellintegrins to immobilized RGDS on cell culture substrates can be reversed spontaneously using mildenvironmental stimulation, such as temperature, without enzymatic or chemical treatment. These findingsare important for control of specific interactions between proteins and cells, and subsequent "on-off"regulation of their function. Furthermore, the method allows serum-free cell culture and trypsin-free cellharvest, essentially removing mammalian-sourced components from the culture process.
C (above the lower critical solutiontemperature, LCST, of the copolymer). Moreover, cells spread on RGDS-immobilized surfaces at 37 Cdetach spontaneously by lowering culture temperature below the LCST as hydrated grafted copolymer chainsdissociate immobilized RGDS from cell integrins. These cell lifting behaviors upon hydration are similar toresults using soluble RGDS in culture as a competitive substitution for immobilized ligands. Binding of cellintegrins to immobilized RGDS on cell culture substrates can be reversed spontaneously using mildenvironmental stimulation, such as temperature, without enzymatic or chemical treatment. These findingsare important for control of specific interactions between proteins and cells, and subsequent "on-off"regulation of their function. Furthermore, the method allows serum-free cell culture and trypsin-free cellharvest, essentially removing mammalian-sourced components from the culture process.