Development of an efficient vitrification method for chondrocyte sheets for clinical application
Introduction: Regenerative therapy using chondrocyte sheets is effective for osteoarthritis. The clinical application of chondrocyte sheet therapy is expected to be further advanced by the use of a feasible cryopreservation technique. Previously, we developed a chondrocyte sheet vitrification method...
| Main Authors: | , , , , , , , |
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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/S2352320420300237 |
| _version_ | 1819078584958976000 |
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| author | Asuka Hayashi Miki Maehara Ayuko Uchikura Hitomi Matsunari Kazuaki Matsumura Suong-Hyu Hyon Masato Sato Hiroshi Nagashima |
| author_facet | Asuka Hayashi Miki Maehara Ayuko Uchikura Hitomi Matsunari Kazuaki Matsumura Suong-Hyu Hyon Masato Sato Hiroshi Nagashima |
| author_sort | Asuka Hayashi |
| collection | DOAJ |
| description | Introduction: Regenerative therapy using chondrocyte sheets is effective for osteoarthritis. The clinical application of chondrocyte sheet therapy is expected to be further advanced by the use of a feasible cryopreservation technique. Previously, we developed a chondrocyte sheet vitrification method; however, it was too complex to be used for routine clinical application. Here, we aimed to develop a prototype method for vitrifying chondrocyte sheets for clinical practice. Methods: We developed a “circulating vitrification bag” as a container to process cell sheets for vitrification in an efficient and sanitary fashion. Moreover, we invented the “vitrification storage box”, which is useful for the vitrification of cell sheets, long-term preservation, and transportation. These devices were used to vitrify rabbit chondrocyte sheets, which were then assessed for their structural characteristics and the viability of the component cells after rewarming. Results: In all cell sheet samples (n = 7) vitrified by the circulating vitrification bag method, the integrity of the sheet structure was maintained, and the cell survival rate was similar to that of non-vitrified samples (91.0 ± 2.9% vs. 90.0 ± 3.0%). Proteoglycan and type II collagen, which are major components of cartilage, were densely and evenly distributed throughout the chondrocyte sheet subjected to vitrification similarly to that observed in the non-vitrified sheet. After long-term storage using the vitrification storage box, the cell sheets maintained normal structure and cell viability (survival rate: 81.2 ± 1.0% vs. 84.3 ± 1.8%) compared to the non-vitrified sheet. Conclusion: Our results indicate that the circulating vitrification bag method is an effective approach for realizing the clinical application of vitrified chondrocyte sheets. The vitrification storage box is also useful for the long-term preservation of vitrified cell sheets, further enhancing the feasibility of the clinical application of cryopreserved chondrocyte sheets. |
| first_indexed | 2024-12-21T19:15:25Z |
| format | Article |
| id | doaj.art-85bcc55e2e52414c81a94cb17be9a7a5 |
| institution | Directory Open Access Journal |
| issn | 2352-3204 |
| language | English |
| last_indexed | 2024-12-21T19:15:25Z |
| publishDate | 2020-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Regenerative Therapy |
| spelling | doaj.art-85bcc55e2e52414c81a94cb17be9a7a52022-12-21T18:53:05ZengElsevierRegenerative Therapy2352-32042020-06-0114215221Development of an efficient vitrification method for chondrocyte sheets for clinical applicationAsuka Hayashi0Miki Maehara1Ayuko Uchikura2Hitomi Matsunari3Kazuaki Matsumura4Suong-Hyu Hyon5Masato Sato6Hiroshi Nagashima7Laboratory of Medical Bioengineering, School of Agriculture, Meiji University, 1-1-1 Higashimita, Tama, Kawasaki 214-8571, JapanDepartment of Orthopaedic Surgery, Surgical Science, Tokai University School of Medicine, 143 Shimokasuya, Isehara, Kanagawa 259-1193, JapanLaboratory of Medical Bioengineering, School of Agriculture, Meiji University, 1-1-1 Higashimita, Tama, Kawasaki 214-8571, JapanLaboratory of Medical Bioengineering, School of Agriculture, Meiji University, 1-1-1 Higashimita, Tama, Kawasaki 214-8571, Japan; Meiji University International Institute for Bio-Resource Research (MUIIBR), 1-1-1 Higashimita, Tama, Kawasaki 214-8571, JapanSchool of Materials Science, Japan Advanced Institute of Science and Technology, 1-1 Asahidai, Nomi, Ishikawa 923-1292 JapanThe Joint Graduate School of Veterinary Medicine, Kagoshima University, Korimoto 1-21-24, Kagoshima 890-8580, JapanDepartment of Orthopaedic Surgery, Surgical Science, Tokai University School of Medicine, 143 Shimokasuya, Isehara, Kanagawa 259-1193, Japan; Meiji University International Institute for Bio-Resource Research (MUIIBR), 1-1-1 Higashimita, Tama, Kawasaki 214-8571, JapanLaboratory of Medical Bioengineering, School of Agriculture, Meiji University, 1-1-1 Higashimita, Tama, Kawasaki 214-8571, Japan; Meiji University International Institute for Bio-Resource Research (MUIIBR), 1-1-1 Higashimita, Tama, Kawasaki 214-8571, Japan; Corresponding author. Laboratory of Medical Bioengineering, School of Agriculture, Meiji University, 1-1-1 Higashimita, Tama, Kawasaki 214-8571, Japan.Introduction: Regenerative therapy using chondrocyte sheets is effective for osteoarthritis. The clinical application of chondrocyte sheet therapy is expected to be further advanced by the use of a feasible cryopreservation technique. Previously, we developed a chondrocyte sheet vitrification method; however, it was too complex to be used for routine clinical application. Here, we aimed to develop a prototype method for vitrifying chondrocyte sheets for clinical practice. Methods: We developed a “circulating vitrification bag” as a container to process cell sheets for vitrification in an efficient and sanitary fashion. Moreover, we invented the “vitrification storage box”, which is useful for the vitrification of cell sheets, long-term preservation, and transportation. These devices were used to vitrify rabbit chondrocyte sheets, which were then assessed for their structural characteristics and the viability of the component cells after rewarming. Results: In all cell sheet samples (n = 7) vitrified by the circulating vitrification bag method, the integrity of the sheet structure was maintained, and the cell survival rate was similar to that of non-vitrified samples (91.0 ± 2.9% vs. 90.0 ± 3.0%). Proteoglycan and type II collagen, which are major components of cartilage, were densely and evenly distributed throughout the chondrocyte sheet subjected to vitrification similarly to that observed in the non-vitrified sheet. After long-term storage using the vitrification storage box, the cell sheets maintained normal structure and cell viability (survival rate: 81.2 ± 1.0% vs. 84.3 ± 1.8%) compared to the non-vitrified sheet. Conclusion: Our results indicate that the circulating vitrification bag method is an effective approach for realizing the clinical application of vitrified chondrocyte sheets. The vitrification storage box is also useful for the long-term preservation of vitrified cell sheets, further enhancing the feasibility of the clinical application of cryopreserved chondrocyte sheets.http://www.sciencedirect.com/science/article/pii/S2352320420300237ChondrocyteCell sheetVitrificationCryopreservationOsteoarthritis |
| spellingShingle | Asuka Hayashi Miki Maehara Ayuko Uchikura Hitomi Matsunari Kazuaki Matsumura Suong-Hyu Hyon Masato Sato Hiroshi Nagashima Development of an efficient vitrification method for chondrocyte sheets for clinical application Regenerative Therapy Chondrocyte Cell sheet Vitrification Cryopreservation Osteoarthritis |
| title | Development of an efficient vitrification method for chondrocyte sheets for clinical application |
| title_full | Development of an efficient vitrification method for chondrocyte sheets for clinical application |
| title_fullStr | Development of an efficient vitrification method for chondrocyte sheets for clinical application |
| title_full_unstemmed | Development of an efficient vitrification method for chondrocyte sheets for clinical application |
| title_short | Development of an efficient vitrification method for chondrocyte sheets for clinical application |
| title_sort | development of an efficient vitrification method for chondrocyte sheets for clinical application |
| topic | Chondrocyte Cell sheet Vitrification Cryopreservation Osteoarthritis |
| url | http://www.sciencedirect.com/science/article/pii/S2352320420300237 |
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