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Fabrication and Validation of Autologous Human Oral Mucosal Epithelial Cell Sheets to Prevent Stenosis after Esophageal Endoscopic Submucosal DissectionTakagi R.a · Yamato M.a · Murakami D.a, d · Kondo M.a, d · Ohki T.a, b · Sasaki R.a, c · Nishida K.e · Namiki H.d · Yamamoto M.b · Okano T.a
aInstitute of Advanced Biomedical Engineering and Science, Tokyo Women’s Medical University and Global Center of Excellence, bDepartment of Surgery, Institute of Gastroenterology, and cDepartment of Oral and Maxillofacial Surgery, Tokyo Women’s Medical University, and dGraduate School of Science and Engineering, Waseda University, Tokyo, and eDepartment of Ophthalmology, Osaka University Medical School, Osaka, Japan Corresponding Author
Masayuki Yamato, PhD
Institute of Advanced Biomedical Engineering and Science Tokyo Women’s Medical University, TWIns, 8-1 Kawada-cho, Shinjuku-ku
Tokyo 162-8666 (Japan)
Tel. +81 3 3353 8112, ext. 66211, E-Mail firstname.lastname@example.org
Objectives: Human oral mucosal epithelial cells derived from 7 healthy volunteer donors were cultured in a clean room in a cell-processing center (CPC) according to good manufacturing practice guidelines. Cell culture and fabricated transplantable epithelial cell sheets were validated for treating ulcers after endoscopic mucosal dissection. Methods: The clonal growth and morphology of the human oral mucosal epithelial cells seeded on temperature-responsive surfaces were observed. During the cultivation, sterilization tests were performed to validate the environment in the CPC. To validate the purity and morphology of fabricated epithelial cell sheets, cell sheets harvested from temperature-responsive surfaces by temperature reduction were examined by histology and flow cytometry. Results: Human oral mucosal epithelial cells were successfully cultured and harvested as continuous cell sheets from temperature-responsive culture inserts without any animal-derived materials. During the cultivations, the sterile environment in the CPC was confirmed. The results of histological and flow cytometry analysis showed the high reproducibility of stratification and the purity of the fabricated human oral mucosal epithelial cell sheets. Conclusions: The method for fabricating epithelial cell sheets shown in this study was suitable for the validation for clinical trials and suggested usability of the fabricated cell sheets.
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