title: Corneal regeneration by transplantation of corneal epithelial cell sheets fabricated with automated cell culture system in rabbit model.
abstract: We have performed clinical applications of cell sheet-based regenerative medicine with human patients in several fields. In order to achieve the mass production of transplantable cell sheets, we have developed automated cell culture systems. Here, we report an automated robotic system utilizing a cell culture vessel, cell cartridge. The cell cartridge had two rooms for epithelial cells and feeder layer cells separating by porous membrane on which a temperature-responsive polymer was covalently immobilized. After pouring cells into this robotic system, cell seeding, medium change, and microscopic examination during culture were automatically performed according to the computer program. Transplantable corneal epithelial cell sheets were successfully fabricated in cell cartridges with this robotic system. Then, fabricated cell sheets were transplanted onto ocular surfaces of rabbit limbal epithelial stem cell deficiency model after 6-h transportation using a portable homothermal container to keep inner temperature at 36 °C. Within one week after transplantation, normal corneal epithelium was successfully regenerated. This automatic cell culture system would be useful for industrialization of tissue-engineered products for regenerative medicine.
authors: Central Research Laboratory, Hitachi, Ltd., 2520 Akanuma, Hiki-gun, Hatoyama-machi, Saitama 350-0395, Japan; Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University, TWIns, 8-1 Kawada-cho, Shinjuku-ku, Tokyo 162-8666, Japan.
authors: Toyoshige
authors: T
authors: Kobayashi
authors: Kazutoshi
authors: K
authors: Kan
authors: Kohji
authors: K
authors: Nishida
authors: Masayuki
authors: M
authors: Yamato
authors: Teruo
authors: T
authors: Okano
mesh_terms: D000818:Animals; D001331:Automation; D018929:Cell Culture Techniques / Q000379:methods*; D003315:Cornea / Q000502:physiology*; D004847:Epithelial Cells / Q000166:cytology* / Q000637:transplantation*; D019573:Epithelium, Corneal / Q000166:cytology*; D006801:Humans; D023421:Models, Animal; D011817:Rabbits; D019651:Plastic Surgery Procedures; D012038:Regeneration; D013696:Temperature; D014186:Transportation
keywords: Automatic cell culture system; Cell cartridge; Cell sheet engineering; Cell sheet transplantation; Cell sheet transportation; Corneal epithelial cell