期刊
BIOMATERIALS
卷 34, 期 17, 页码 4214-4222出版社
ELSEVIER SCI LTD
DOI: 10.1016/j.biomaterials.2013.02.056
关键词
Poly(N-isopropylacrylamide); Affinity; Thermoresponsive cell culture surface; Heparin; Basic fibroblast growth factor; Cell sheet
资金
- Global COE Program, Multidisciplinary Education and Research Center for Regenerative Medicine (MERCREM) from the Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan
- Creation of Innovation Centers for Advanced Interdisciplinary Research Areas Program in the Project for Developing Innovation Systems Cell Sheet Tissue Engineering Center (CSTEC) from MEXT of Japan
- MEXT of Japan [23106009, 2305]
- Japan Society for the Promotion of Science (JSPS) [10J57081]
- Grants-in-Aid for Scientific Research [10J57081] Funding Source: KAKEN
Heparin-functionalized poly(N-isopropylacrylamide-co-2-carboxyisopropylacrylamide) [P(IPAAm-co-CIPAAm)] grafted surface was designed for the switching of cell growth/detachment, achieved by the regulation of affinity binding between basic fibroblast growth factor (bFGF) and immobilized heparin through the temperature-dependent conformational change of grafted P(IPAAm-co-CIPAAm) chains. At 37 degrees C, bFGF-bound heparin-thermoresponsive surfaces were able to hold the two- to three-fold number of mouse fibroblast (NIH/3T3) cells than both bFGF-physisorbed surface and PIPAAm surface with soluble bFGF after a 3-day cultivation. Bound bFGF via heparin on shrunken grafted P(IPAAm-co-CIPAAm) chains at 37 degrees C was able to reinforce the formation and stabilization of bFGF-FGF receptor complex, although the activity of physisorbed bFGF on PIPAAm-grafted surfaces was decreased by non-specific and randomly oriented adsorption. At 20 degrees C, the cultured NIH/3T3 cell sheet with bFGF detached from heparin-functionalized thermoresponsive surface. The release of bFGF from the surfaces was induced by reducing the affinity binding between bFGF and immobilized-heparin due to increasing the mobility of the swollen grafted P(IPAAm-co-CIPAAm) chains. Therefore, heparin-functionalized thermoresponsive surface was able to enhance cell proliferation, and confluent cells detached themselves as a contiguous cell sheet due to switching cell growth by changing temperature. A cell culture system using this surface is useful for rapid cell sheet fabrication and manipulation. (C) 2013 Elsevier Ltd. All rights reserved.
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