Elsevier

Progress in Polymer Science

Volume 32, Issues 8–9, August–September 2007, Pages 1123-1133
Progress in Polymer Science

Temperature-responsive cell culture surfaces for regenerative medicine with cell sheet engineering

https://doi.org/10.1016/j.progpolymsci.2007.06.002Get rights and content

Abstract

We have developed a novel strategy for regenerative medicine to recover tissue functions by using temperature-responsive cell culture surfaces. To overcome the limits of conventional methods such as the use of single-cell suspension injection and the use of biodegradable polymer scaffolds, we have applied transplantable cell sheets fabricated with temperature-responsive culture surfaces for cell delivery. Here, we summarize the development of the next generation of temperature-responsive cell culture surfaces using modifications of the polymer coating to allow for rapid cell sheet recovery; improved culture conditions, such that animal-derived products can be eliminated; and altered phase transition temperatures to produce patterned cell sheets. These designs allow for the reconstruction of more complex tissues to potentially treat a wide range of diseases.

Section snippets

Conventional methods for regenerative medicine

Regenerative medicine has been expected to achieve functional recovery of damaged tissues. Although promising results have been obtained with conventional cell therapies, there still remain numerous problems to regenerate functional tissues. The most widely used method for cell delivery is direct injection of single-cell suspensions. Clinical success has been reported in the treatment of Parkinson's disease [1], [2] and Huntington's disease [3] by the injection of neural cells obtained from

Rapid cell sheet detachment from poly(N-isopropylacrylamide)-grafted porous cell culture membranes

Cell detachment from temperature-responsive culture surfaces after temperature reduction, is driven by cell traction forces exerted by the cytoskeleton [47]. The amount and properties of ECM deposited by cultured cells are also key factors in cell detachment from temperature-responsive surfaces. Since the ECM varies significantly among cell types, effective cell detachment cannot be achieved or requires prolonged time for complete cell harvest, with cell sheets of some cell types [48].

In order

Copolymerization of 2-carboxyisopropylacrylamide with N-isopropylacrylamide accelerates cell sheet detachment

Acrylic acid (AAc) has been often utilized to introduce reactive carboxyl groups to the temperature-responsive polymer, PIPAAm. However, AAc introduction shifts the copolymer phase transition temperatures higher and dampens the steep homopolymer phase transition with increasing AAc content. We previously synthesized 2-carboxyisopropylacrylamide (CIPAAm) having both a similar side chain structure to IPAAm and a functional carboxylate group in order to overcome these shortcomings [54]. AAc

Control of phase transition temperature

The hydrophobic monomer, n-butyl methacrylate (BMA) has been incorporated into temperature-responsive PIPAAm to lower PIPAAm phase transition temperatures necessary for systematically regulating cell adhesion on and detachment from culture dishes at controlled temperatures. Poly(IPAAm-co-BMA)-grafted dishes were prepared by electron beam irradiation methods, with systematic changes in the BMA content of the feed monomer. Copolymer-grafted surfaces showed decreased grafted polymer transition

Conclusion

We have developed a novel strategy to fabricate functional tissues in regenerative medicine by focusing on transplantable cell sheets. Temperature-responsive culture surfaces are unique tools to contribute to new clinical achievements. Further modifications can be expected to realize more complex tissue structures and functions by cell sheet engineering.

Acknowledgments

We appreciate the continued useful comments and technical criticism from Prof. D.W. Grainger (University of Utah, USA). This work was supported in part by Grants-in-Aid for Scientific Research by Ministry of Education, Culture, Sports, Science and Technology of Japan.

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    1

    Current address: Department of Materials Science and Technology, Tokyo University of Science, 2641 Yamazaki, Noda-shi, Chiba 278-8510, Japan.

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