Fabrication of tissue-engineered cell sheets by automated cell culture equipment

利用自动化细胞培养设备制作组织工程细胞片

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作者:Ayako Nishimura, Ryota Nakajima, Ryo Takagi, Guangbin Zhou, Daisuke Suzuki, Masaharu Kiyama, Takayuki Nozaki, Toshiyuki Owaki, Tomomi Takahara, Shigeru Nagai, Taku Nakamura, Masakazu Sugaya, Koichi Terada, Yumiko Igarashi, Hiroko Hanzawa, Teruo Okano, Tatsuya Shimizu, Masayuki Yamato, Shizu Takeda

Abstract

Most cells for regenerative medicine are currently cultured manually. In order to promote the widespread use of regenerative medicine, it will be necessary to develop automated culture techniques so that cells can be produced in greater quantities at lower cost and with more stable quality. In the field of regenerative medicine technology, cell sheet therapy is an effective tissue engineering technique whereby cells can be grafted by attaching them to a target site. We have developed automated cell culture equipment to promote the use of this cell sheet regenerative treatment. This equipment features a fully closed culture vessel and circuit system that avoids contamination with bacteria and the like from the external environment, and it was designed to allow 10 cell sheets to be simultaneously cultured in parallel. We used this equipment to fabricate 50 sheets of human oral mucosal epithelial cells in five automated culture tests in this trial. By analyzing these sheets, we confirmed that 49 of the 50 sheets satisfied the quality standards of clinical research. To compare the characteristics of automatically fabricated cell sheets with those of manually fabricated cell sheets, we performed histological analyses using immunostaining and transmission electron microscopy. The results confirmed that cell sheets fabricated with the automated cell culture are differentiated in the same way as cultures fabricated manually.

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