A stable protocol for the fabrication of transplantable human oral mucosal epithelial cell sheets for clinical application
Introduction: Cultured stratified epithelial cell sheets have been clinically utilized as transplantable grafts for the regeneration of epithelial tissues, such as the esophagus, cornea, skin, and intraoral cavity. These cell sheets are expected to gain widespread use as regenerative medicine produc...
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| Format: | Article |
| Language: | English |
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Elsevier
2020-06-01
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| Series: | Regenerative Therapy |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2352320419301579 |
| _version_ | 1828862784234848256 |
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| author | Yoshiyuki Kasai Ryo Takagi Shinichiro Kobayashi Toshiyuki Owaki Naoyuki Yamaguchi Hiroko Fukuda Yusuke Sakai Yoshinori Sumita Nobuo Kanai Hajime Isomoto Kengo Kanetaka Takeshi Ohki Izumi Asahina Kazuhiro Nagai Kazuhiko Nakao Naoya Takeda Teruo Okano Susumu Eguchi Masayuki Yamato |
| author_facet | Yoshiyuki Kasai Ryo Takagi Shinichiro Kobayashi Toshiyuki Owaki Naoyuki Yamaguchi Hiroko Fukuda Yusuke Sakai Yoshinori Sumita Nobuo Kanai Hajime Isomoto Kengo Kanetaka Takeshi Ohki Izumi Asahina Kazuhiro Nagai Kazuhiko Nakao Naoya Takeda Teruo Okano Susumu Eguchi Masayuki Yamato |
| author_sort | Yoshiyuki Kasai |
| collection | DOAJ |
| description | Introduction: Cultured stratified epithelial cell sheets have been clinically utilized as transplantable grafts for the regeneration of epithelial tissues, such as the esophagus, cornea, skin, and intraoral cavity. These cell sheets are expected to gain widespread use as regenerative medicine products and save many patients. For this purpose, establishing and disseminating the stale protocol of fabricating the cell sheet is crucial. The fabrication of cultured stratified epithelial cell sheets consists of many important steps, and since the patients’ epithelial cell conditions vary widely and are sometimes unstable, the qualities of the epithelial cell grafts are likewise potentially unstable. Therefore, in this paper, we report the stable protocol for fabrication of the transplantable cell sheet particularly from patient-derived oral mucosal tissues. Methods: Serum extracted from blood and buccal mucosal tissue were collected in Nagasaki University and transported to Tokyo Women's Medical University. Oral mucosal epithelial cells were collected by minimum trypsin method, and this treatment was studied whether to be a critical procedure. After 14 days cultivation, cultured cells were examined whether to be transplantable as cell sheets. Results: We successfully transported buccal mucosal tissue and serum without damage and contamination. Oral mucosal epithelial cells were collected with high viability by minimum trypsin method. Finally, we succeeded to stably fabricate oral mucosal epithelial cell sheets in all 10 patients. Conclusions: We established a stable protocol for the fabrication of human oral mucosal epithelial cell sheets and their transportation in clinical settings in this study. These methodologies could also be basis for transplantation therapy using cultured cell sheets of various types other than oral mucosal epithelial cell and will contribute largely to the future development of regenerative medicine. |
| first_indexed | 2024-12-13T03:30:54Z |
| format | Article |
| id | doaj.art-72f6a459f6624a6dad23539c231293c6 |
| institution | Directory Open Access Journal |
| issn | 2352-3204 |
| language | English |
| last_indexed | 2024-12-13T03:30:54Z |
| publishDate | 2020-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Regenerative Therapy |
| spelling | doaj.art-72f6a459f6624a6dad23539c231293c62022-12-22T00:01:10ZengElsevierRegenerative Therapy2352-32042020-06-01148794A stable protocol for the fabrication of transplantable human oral mucosal epithelial cell sheets for clinical applicationYoshiyuki Kasai0Ryo Takagi1Shinichiro Kobayashi2Toshiyuki Owaki3Naoyuki Yamaguchi4Hiroko Fukuda5Yusuke Sakai6Yoshinori Sumita7Nobuo Kanai8Hajime Isomoto9Kengo Kanetaka10Takeshi Ohki11Izumi Asahina12Kazuhiro Nagai13Kazuhiko Nakao14Naoya Takeda15Teruo Okano16Susumu Eguchi17Masayuki Yamato18Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University (TWIns), 8-1 Kawada-Cho, Shinjuku-ku, Tokyo, 162-8666, Japan; Department of Life Science and Medical Bioscience, Graduate School of Advanced Science and Engineering, Waseda University (TWIns), 2-2 Wakamatsu-Cho, Shinjuku-ku, Tokyo, 162-8480, JapanInstitute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University (TWIns), 8-1 Kawada-Cho, Shinjuku-ku, Tokyo, 162-8666, JapanInstitute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University (TWIns), 8-1 Kawada-Cho, Shinjuku-ku, Tokyo, 162-8666, Japan; Department of Surgery, Nagasaki University Graduate School of Biomedical Sciences, 1-7-1 Sakamoto, Nagasaki-Shi, Nagasaki, 852-8501, JapanInstitute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University (TWIns), 8-1 Kawada-Cho, Shinjuku-ku, Tokyo, 162-8666, JapanDepartment of Gastroenterology and Hepatology, Nagasaki University Hospital, 1-7-1 Sakamoto, Nagasaki-Shi, Nagasaki, 852-8501, JapanDepartment of Gastroenterology and Hepatology, Nagasaki University Hospital, 1-7-1 Sakamoto, Nagasaki-Shi, Nagasaki, 852-8501, JapanDepartment of Surgery, Nagasaki University Graduate School of Biomedical Sciences, 1-7-1 Sakamoto, Nagasaki-Shi, Nagasaki, 852-8501, JapanDepartment of Regenerative Oral Surgery, Nagasaki University Graduate School of Biomedical Sciences, 1-7-1 Sakamoto, Nagasaki-Shi, Nagasaki, 852-8501, JapanInstitute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University (TWIns), 8-1 Kawada-Cho, Shinjuku-ku, Tokyo, 162-8666, JapanDepartment of Gastroenterology and Hepatology, Nagasaki University Hospital, 1-7-1 Sakamoto, Nagasaki-Shi, Nagasaki, 852-8501, JapanDepartment of Surgery, Nagasaki University Graduate School of Biomedical Sciences, 1-7-1 Sakamoto, Nagasaki-Shi, Nagasaki, 852-8501, JapanInstitute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University (TWIns), 8-1 Kawada-Cho, Shinjuku-ku, Tokyo, 162-8666, JapanDepartment of Regenerative Oral Surgery, Nagasaki University Graduate School of Biomedical Sciences, 1-7-1 Sakamoto, Nagasaki-Shi, Nagasaki, 852-8501, JapanTransfusion and Cell Therapy Unit, Nagasaki University Hospital, 1-7-1 Sakamoto, Nagasaki-Shi, Nagasaki, 852-8501, JapanDepartment of Gastroenterology and Hepatology, Nagasaki University Hospital, 1-7-1 Sakamoto, Nagasaki-Shi, Nagasaki, 852-8501, JapanDepartment of Life Science and Medical Bioscience, Graduate School of Advanced Science and Engineering, Waseda University (TWIns), 2-2 Wakamatsu-Cho, Shinjuku-ku, Tokyo, 162-8480, JapanInstitute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University (TWIns), 8-1 Kawada-Cho, Shinjuku-ku, Tokyo, 162-8666, JapanDepartment of Surgery, Nagasaki University Graduate School of Biomedical Sciences, 1-7-1 Sakamoto, Nagasaki-Shi, Nagasaki, 852-8501, JapanInstitute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University (TWIns), 8-1 Kawada-Cho, Shinjuku-ku, Tokyo, 162-8666, Japan; Corresponding author. Fax: +81-3-3359-6046.Introduction: Cultured stratified epithelial cell sheets have been clinically utilized as transplantable grafts for the regeneration of epithelial tissues, such as the esophagus, cornea, skin, and intraoral cavity. These cell sheets are expected to gain widespread use as regenerative medicine products and save many patients. For this purpose, establishing and disseminating the stale protocol of fabricating the cell sheet is crucial. The fabrication of cultured stratified epithelial cell sheets consists of many important steps, and since the patients’ epithelial cell conditions vary widely and are sometimes unstable, the qualities of the epithelial cell grafts are likewise potentially unstable. Therefore, in this paper, we report the stable protocol for fabrication of the transplantable cell sheet particularly from patient-derived oral mucosal tissues. Methods: Serum extracted from blood and buccal mucosal tissue were collected in Nagasaki University and transported to Tokyo Women's Medical University. Oral mucosal epithelial cells were collected by minimum trypsin method, and this treatment was studied whether to be a critical procedure. After 14 days cultivation, cultured cells were examined whether to be transplantable as cell sheets. Results: We successfully transported buccal mucosal tissue and serum without damage and contamination. Oral mucosal epithelial cells were collected with high viability by minimum trypsin method. Finally, we succeeded to stably fabricate oral mucosal epithelial cell sheets in all 10 patients. Conclusions: We established a stable protocol for the fabrication of human oral mucosal epithelial cell sheets and their transportation in clinical settings in this study. These methodologies could also be basis for transplantation therapy using cultured cell sheets of various types other than oral mucosal epithelial cell and will contribute largely to the future development of regenerative medicine.http://www.sciencedirect.com/science/article/pii/S2352320419301579Primary cell cultureMinimum trypsinizationEpithelial cell sheetClinical application |
| spellingShingle | Yoshiyuki Kasai Ryo Takagi Shinichiro Kobayashi Toshiyuki Owaki Naoyuki Yamaguchi Hiroko Fukuda Yusuke Sakai Yoshinori Sumita Nobuo Kanai Hajime Isomoto Kengo Kanetaka Takeshi Ohki Izumi Asahina Kazuhiro Nagai Kazuhiko Nakao Naoya Takeda Teruo Okano Susumu Eguchi Masayuki Yamato A stable protocol for the fabrication of transplantable human oral mucosal epithelial cell sheets for clinical application Regenerative Therapy Primary cell culture Minimum trypsinization Epithelial cell sheet Clinical application |
| title | A stable protocol for the fabrication of transplantable human oral mucosal epithelial cell sheets for clinical application |
| title_full | A stable protocol for the fabrication of transplantable human oral mucosal epithelial cell sheets for clinical application |
| title_fullStr | A stable protocol for the fabrication of transplantable human oral mucosal epithelial cell sheets for clinical application |
| title_full_unstemmed | A stable protocol for the fabrication of transplantable human oral mucosal epithelial cell sheets for clinical application |
| title_short | A stable protocol for the fabrication of transplantable human oral mucosal epithelial cell sheets for clinical application |
| title_sort | stable protocol for the fabrication of transplantable human oral mucosal epithelial cell sheets for clinical application |
| topic | Primary cell culture Minimum trypsinization Epithelial cell sheet Clinical application |
| url | http://www.sciencedirect.com/science/article/pii/S2352320419301579 |
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