Fabrication of functional three-dimensional tissues by stacking cell sheets in vitro

Yuji Haraguchi, Tatsuya Shimizu, Tadashi Sasagawa, Hidekazu Sekine, Katsuhisa Sakaguchi, Tetsutaro Kikuchi, Waki Sekine, Sachiko Sekiya, Masayuki Yamato, Mitsuo Umezu, Teruo Okano*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

305 Citations (Scopus)


The fabrication of 3D tissues retaining the original functions of tissues/organs in vitro is crucial for optimal tissue engineering and regenerative medicine. The fabrication of 3D tissues also contributes to the establishment of in vitro tissue/organ models for drug screening. Our laboratory has developed a fabrication system for functional 3D tissues by stacking cell sheets of confluent cultured cells detached from a temperature-responsive culture dish. Here we describe the protocols for the fabrication of 3D tissues by cell sheet engineering. Three-dimensional cardiac tissues fabricated by stacking cardiac cell sheets pulsate spontaneously, synchronously and macroscopically. Via this protocol, it is also possible to fabricate other tissues, such as 3D tissue including capillary-like prevascular networks, from endothelial cells sandwiched between layered cell sheets. Cell sheet stacking technology promises to provide in vitro tissue/organ models and more effective therapies for curing tissue/organ failures.

Original languageEnglish
Pages (from-to)850-858
Number of pages9
JournalNature protocols
Issue number5
Publication statusPublished - 2012 May

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)


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