Selective Cell Adhesion and Detachment on Antibody-Immobilized Thermoresponsive Surfaces by Temperature Changes
Jun Kobayashi*, Masanori Nishi*,**, Yoshikatsu Akiyama*,
Masayuki Yamato*, Hirofumi Yajima**, and Teruo Okano*
*Institute of Advanced Biomedical Engineering and Science, Tokyo Women’s Medical University (TWIns), 8-1 Kawadacho, Shinjuku, Tokyo 162-8666, Japan
**Department of Applied Chemistry, Faculty of Science, Tokyo University of Science, 1-3 Kagurazaka, Shinjuku, Tokyo 162-8601, Japan
Anti-CD90 antibody-immobilized thermoresponsive (AIT) surfaces were prepared for obtaining temperature-triggered switching of the selective adhesion and detachment of CD90-expressed cells. Thymic carcinoma cells (Ty-82) expressing CD90 molecules on the cellular surface were unable to adhere to isotype AIT surfaces and aggregated. In contrast, Ty-82 cells selectively adhered to anti-CD90 AIT surfaces at 37°C. These results indicate that Ty-82 cells adhered to CD90 antibody-immobilized surfaces through affinity interaction, not through nonspecific interactions when grafted thermoresponsive polymer chains shrunk at 37°C. Adhered cells were detached from surfaces by lowering temperature to 20°C with pipetting. Although affinity interaction between cells and immobilized antibodies was decreased by the dynamic swelling of grafted thermoresponsive polymer chains by lowering temperature to 20°C, the application of additional force such as pipetting was required to completely detach adhered cells. Through temperature-induced changes in affinity interaction, the condensation of CD90-positive Ty-82 cells was carried out by using anti-CD90 AIT surfaces. AIT surfaces for regulating selective cell adhesion and detachment were then successfully prepared. A novel bioassembler technology using AIT surfaces could thus be useful for temperature-dependent selective cell adhesion/detachment such as cell separation.
Masayuki Yamato, Hirofumi Yajima, and Teruo Okano, “Selective Cell Adhesion and Detachment on Antibody-Immobilized Thermoresponsive Surfaces by Temperature Changes,” J. Robot. Mechatron., Vol.25, No.4, pp. 637-643, 2013.
-  R. Langer and J. P. Vacanti, “Tissue Engineering,” Science, Vol.260, No.5110, pp. 920-926, 1993.
-  M. Yamato and T. Okano, “Cell Sheet Engineering,” Mater. Today, Vol.7, No.5, pp. 42-47, 2004.
-  N. Yamada, T. Okano et al., “Thermo-responsive Polymeric Surfaces; Control of Attachment and Detachment of Cultured Cells,” Makromol. Chem. Rapid. Commun., Vol.11, No.11, pp. 571-576, 1990.
-  M. Heskins and J. E. Guillet, “Solution Properties of Poly(Nisopropylacrylamide),” J. Macromol. Sci. A Chem., Vol.2, No.8, pp. 1441-1455, 1968.
-  J. Kobayashi and T. Okano, “Fabrication of a Thermoresponsive Cell Culture Dish: a Key Technology for Cell Sheet Tissue Engineering,” J. Sci. Technol. Adv. Mater., Vol.11, p. 014111, 2010.
-  K. Matsuura, M. Wada et al., “Creation of Human Cardiac Cell Sheets Using Pluripotent Stem Cells,” Biochem. Biophys. Res. Commun., Vol.425, No.2, pp. 321-327, 2012.
-  I.Minami, K. Yamada et at., “A Small Molecule that Promotes Cardiac Differentiation of Human Pluripotent Stem Cells under Defined, Cytokine- and Xeno-free Conditions,” Cell Reports, Vol.2, No.5, pp. 1448-1460, 2012.
-  D. Mattanovich and N. Borth, “Applications of Cell Sorting in Biotechnology,” Microbial Cell. Factories, Vol.5, p. 12, 2006.
-  T. A. Rege and J. S. Hagood, “Thy-1 as a Regulator of Cell-cell and Cell-matrix Interactions in Axon Regeneration, Apoptosis, Adhesion, Migration, Cancer, and Fibrosis,” FASEB J., Vol.20, No,8, pp. 1045-1054, 2006.
-  T. Aoyagi, M. Ebara et al., “Novel Bifunctional Polymer with Reactivity and Temperature Sensitivity,” J. Biomater. Sci. Polym. Ed., Vol.11, No.1, pp. 101-110, 2000.
-  M. Nishi, J. Kobayashi et al., “The Use of Biotin-avidin Binding to Facilitate Biomodification of Thermoresponsive Culture Surfaces,” Biomaterials, Vol.28, No.36, pp. 5471-5476, 2007.
-  A. Tamura, J. Kobayashi et al., “Temperature-responsive Poly(Nisopropylacrylamide)-grafted Microcarriers for Large-scale Noninvasive Harvest of Anchorage-dependent Cells,” Biomaterials, Vol.33, No.15, pp. 3803-3812, 2012.
This article is published under a Creative Commons Attribution-NoDerivatives 4.0 International License.
Copyright© 2013 by Fuji Technology Press Ltd. and Japan Society of Mechanical Engineers. All right reserved.