Surface design of antibody-immobilized thermoresponsive cell culture dishes for recovering intact cells by low-temperature treatment
Article first published online: 20 DEC 2013
© 2013 Wiley Periodicals, Inc.
Journal of Biomedical Materials Research Part A
Volume 102, Issue 11, pages 3883–3893, November 2014
How to Cite
How to cite this article: 2014. Surface design of antibody-immobilized thermoresponsive cell culture dishes for recovering intact cells by low-temperature treatment. J Biomed Mater Res Part A 2014:102A:3883–3893., , , , , , , , , , .
- Issue published online: 25 SEP 2014
- Article first published online: 20 DEC 2013
- Accepted manuscript online: 12 DEC 2013 12:46AM EST
- Manuscript Accepted: 9 DEC 2013
- Manuscript Revised: 21 NOV 2013
- Manuscript Received: 2 OCT 2013
- Ministry of Education, Culture, Sports, Science and Technology, Japan (MEXT), Creation of Innovation Centers for Advanced Interdisciplinary Research Areas Program in the Project for Developing Innovation Systems “Cell Sheet Tissue Engineering Center (CSTEC)”
- The Japan Society for the Promotion of Science (JSPS), Grant-in-Aid for Scientific Research on Innovative Areas “Bio Assembler”. Grant Number: 23106009
- MEXT, Global COE Program, Multidisciplinary Education and Research Center for Regenerative Medicine (MERCREM)
- JSPS, Grant-in-Aid for Young Scientists (B). Grant Number: 25750179
- thermoresponsive cell culture surface;
- cell sheet;
- adipose tissue-derived cell
Antibody-immobilized thermoresponsive poly(N-isopropylacrylamide-co-2-carboxyisopropylacrylamide) [poly(IPAAm-co-CIPAAm)]-grafted cell culture surfaces were designed to enhance both the initial adhesion of weakly adhering cells and the ability of cells to detach in response to low temperature through the regulation of affinity binding between immobilized antibodies and antigens on the cellular surface. Ty-82 cells and neonatal normal human dermal fibroblasts (NHDFs), which express CD90 on the cell surface, adhered to anti-CD90 antibody-immobilized thermoresponsive surfaces at 37°C, a condition at which the grafted thermoresponsive polymer chains shrank. Adherent Ty-82 cells were detached from the surfaces by lowering the temperature to 20°C and applying external forces, such as pipetting, whereas cultured NHDF sheets spontaneously detached themselves from the surface in response to reduced temperature alone. When the temperature was decreased to 20°C, the swelling of grafted thermoresponsive polymer chains weakened the affinity binding between immobilized antibody and antigen on the cells due to the increasing steric hindrance of the polymer chains around the antigen-recognition site of the immobilized antibodies. No contamination was detected on cells harvested from covalently immobilized antibodies on the culture surfaces by low-temperature treatment, whereas a carryover of the antibody and avidin from the avidin-biotin binding surface was observed. Furthermore, the initial adhesion of adipose tissue-derived cells, which adhere weakly to PIPAAm-grafted surfaces, was enhanced on the antibody-immobilized thermoresponsive surfaces. © 2013 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 102A: 3883–3893, 2014.