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Reproducible subcutaneous transplantation of cell sheets into recipient mice

A Retraction to this article was published on 25 February 2016

This article has been updated


Perfecting tissue engineering and cell sheet transplantation is an important step toward realizing regenerative medicine and is a growing area of research. Before being applied to clinical settings, it is important that these approaches are evaluated in vivo. Here we provide a detailed protocol for handling thin cell sheets, for a simple and highly reproducible subcutaneous transplantation of cell sheets into mice, and for the histological examination of regenerated tissues. Various aspects of transplants can be assessed, such as maintenance, differentiation and proliferation. An emphasis is placed on surgical precision and reproducibility. The resulting consistency between surgeries helps minimize artifacts from surgical variation and therefore enables researchers to not only observe and compare the interactions between host tissues but also to compare transplants among different host animals. A single transplantation can be carried out within 10 min.

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Figure 1: Preparation of transplantable cell sheets.
Figure 2: Subcutaneous transplantation.
Figure 3: Tissue preparation for histology.
Figure 4: Histology of regenerated epidermal tissue in subcutaneous space.
Figure 5: Quantitative examination of inflammatory cell reaction.

Change history

  • 13 January 2016

     Masayuki Yamato, Satoshi Tsuneda and Teruo Okano would like to retract this protocol after concerns were raised by the community about some of the figures. Specifically, concerns were raised that the fourth graph in Figure 5a and the first graph in Figure 5b look very similar, and some of the error bars look unevenly positioned. Masayuki Yamato, Satoshi Tsuneda and Teruo Okano have been unable to locate some of the raw data to verify these figures and are no longer confident in the paper's results. Given that these results are key to demonstrating the reliability and reproducibility of the protocol, these authors wish to retract the protocol, and they sincerely apologize for the adverse consequences that may have resulted from its publication. Haruko Obokata could not be reached by the journal for comment on the retraction.


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This study was partially supported by: Grants-in-Aid for JSPS Fellows; the Formation of Innovation Center for Fusion of Advanced Technologies in the Special Coordination Funds for Promoting Science and Technology; and the Global COE program. All these grants are from from the Ministry of Education, Culture, Sports, Science and Technology, Japan.

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Authors and Affiliations



H.O. designed and conducted the experiments, analyzed the data and wrote the paper. M.Y. designed experiments, analyzed data and wrote the paper. S.T. and T.O. supervised the project.

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Correspondence to Teruo Okano.

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The authors declare no competing financial interests.

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Obokata, H., Yamato, M., Tsuneda, S. et al. Reproducible subcutaneous transplantation of cell sheets into recipient mice. Nat Protoc 6, 1053–1059 (2011).

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