Stable preparation of in vivo transplantable periodontal ligament-derived mesenchymal stem cell sheets in thermoresponsive culture dishes with tunable cell detachability

Tissue engineering plays a pivotal role in the advancement of regenerative medicine. Thermoresponsive culture dishes, coated with specialized polymers that control cell adhesion through temperature fluctuations, enable the processing of cells into sheets for medical applications while maintaining th...

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Main Authors: Kazuki Morita, Masamichi Nakayama, Jiacheng Wang, Satoru Onizuka, Masahiro Hatasa, Yujin Ohsugi, Yosuke Tsuchiya, Hiromi Niimi, Anhao Liu, Hideaki Sakai, Teruo Okano, Takanori Iwata
Format: Article
Language:English
Published: Elsevier 2025-03-01
Series:Regenerative Therapy
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Online Access:http://www.sciencedirect.com/science/article/pii/S2352320425000045
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author Kazuki Morita
Masamichi Nakayama
Jiacheng Wang
Satoru Onizuka
Masahiro Hatasa
Yujin Ohsugi
Yosuke Tsuchiya
Hiromi Niimi
Anhao Liu
Hideaki Sakai
Teruo Okano
Takanori Iwata
author_facet Kazuki Morita
Masamichi Nakayama
Jiacheng Wang
Satoru Onizuka
Masahiro Hatasa
Yujin Ohsugi
Yosuke Tsuchiya
Hiromi Niimi
Anhao Liu
Hideaki Sakai
Teruo Okano
Takanori Iwata
author_sort Kazuki Morita
collection DOAJ
description Tissue engineering plays a pivotal role in the advancement of regenerative medicine. Thermoresponsive culture dishes, coated with specialized polymers that control cell adhesion through temperature fluctuations, enable the processing of cells into sheets for medical applications while maintaining their intact state. Cell sheets prepared using these culture dishes have been incorporated into several commercial pharmaceutical products. However, controlling the detachability of cell sheets using conventional thermoresponsive culture dishes remains a challenge, and often leads to unexpected detachment during cultivation. In this study, we developed a thermoresponsive culture dish with tunable cell detachability using a thermoresponsive block copolymer, poly(butyl methacrylate)-b-poly(N-isopropylacrylamide) (PBMA-PIPAAm), which is a specialized polymer that allows precise control of the amount of surface-immobilized polymer and polymer layer thickness. Culturing periodontal ligament-derived mesenchymal stem cells on these dishes demonstrated fully tunable detachability without compromising cell properties compared to conventional thermoresponsive dishes (UpCell®). Thermoresponsive PBMA-PIPAAm-coated culture dishes enable the complete on-demand detachment of transplantable cell sheets, thereby avoiding unexpected detachment that may increase production costs and reduce technical hurdles in the manufacturing process. The PBMA-PIPAAm coating method has the potential to contribute to biomedical and clinical applications of mesenchymal stem cell sheets.
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spelling doaj-art-95279d53e96847ffad1ccc3131000d212025-08-20T02:46:59ZengElsevierRegenerative Therapy2352-32042025-03-012834535710.1016/j.reth.2025.01.004Stable preparation of in vivo transplantable periodontal ligament-derived mesenchymal stem cell sheets in thermoresponsive culture dishes with tunable cell detachabilityKazuki Morita0Masamichi Nakayama1Jiacheng Wang2Satoru Onizuka3Masahiro Hatasa4Yujin Ohsugi5Yosuke Tsuchiya6Hiromi Niimi7Anhao Liu8Hideaki Sakai9Teruo Okano10Takanori Iwata11Department of Periodontology, Graduate School of Medical and Dental Sciences, Institute of Science Tokyo (Science Tokyo), 1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8510, JapanInstitute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University, 8-1 Kawadacho, Shinjuku-ku, Tokyo 162-8666, JapanDepartment of Periodontology, Graduate School of Medical and Dental Sciences, Institute of Science Tokyo (Science Tokyo), 1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8510, JapanDivision of Periodontology, Department of Oral Function, Kyushu Dental University, 2-6-1 Manazuru, Kokurakita-ku, Kitakyushu, Fukuoka, 803-8580, JapanDepartment of Periodontology, Graduate School of Medical and Dental Sciences, Institute of Science Tokyo (Science Tokyo), 1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8510, Japan; Liver Center, Saga University Hospital, Saga, Saga, JapanSection of Vascular Cell Biology, Joslin Diabetes Center, Harvard Medical School, One Joslin Place, Boston, MA, 02215, USA; Department of Oral Biology, Graduate School of Medical and Dental Sciences, Institute of Science Tokyo (Science Tokyo), 1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8510, Japan; Oral Science Center, Institute of Science Tokyo (Science Tokyo), 1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8510, JapanDepartment of Periodontology, Graduate School of Medical and Dental Sciences, Institute of Science Tokyo (Science Tokyo), 1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8510, JapanDepartment of Lifetime Oral Health Care Science, Graduate School of Medical and Dental Sciences, Institute of Science Tokyo (Science Tokyo), 1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8510, JapanDepartment of Periodontology, Graduate School of Medical and Dental Sciences, Institute of Science Tokyo (Science Tokyo), 1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8510, JapanOrganization of Cell Sheet Tissue Engineering Regenerative Medicine Initiatives (CSTERM), Ark Mori Building 36F, 1-12-32 Akasaka, Minato-ku, Tokyo 107-6036, JapanInstitute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University, 8-1 Kawadacho, Shinjuku-ku, Tokyo 162-8666, Japan; Organization of Cell Sheet Tissue Engineering Regenerative Medicine Initiatives (CSTERM), Ark Mori Building 36F, 1-12-32 Akasaka, Minato-ku, Tokyo 107-6036, JapanDepartment of Periodontology, Graduate School of Medical and Dental Sciences, Institute of Science Tokyo (Science Tokyo), 1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8510, Japan; Corresponding author.Tissue engineering plays a pivotal role in the advancement of regenerative medicine. Thermoresponsive culture dishes, coated with specialized polymers that control cell adhesion through temperature fluctuations, enable the processing of cells into sheets for medical applications while maintaining their intact state. Cell sheets prepared using these culture dishes have been incorporated into several commercial pharmaceutical products. However, controlling the detachability of cell sheets using conventional thermoresponsive culture dishes remains a challenge, and often leads to unexpected detachment during cultivation. In this study, we developed a thermoresponsive culture dish with tunable cell detachability using a thermoresponsive block copolymer, poly(butyl methacrylate)-b-poly(N-isopropylacrylamide) (PBMA-PIPAAm), which is a specialized polymer that allows precise control of the amount of surface-immobilized polymer and polymer layer thickness. Culturing periodontal ligament-derived mesenchymal stem cells on these dishes demonstrated fully tunable detachability without compromising cell properties compared to conventional thermoresponsive dishes (UpCell®). Thermoresponsive PBMA-PIPAAm-coated culture dishes enable the complete on-demand detachment of transplantable cell sheets, thereby avoiding unexpected detachment that may increase production costs and reduce technical hurdles in the manufacturing process. The PBMA-PIPAAm coating method has the potential to contribute to biomedical and clinical applications of mesenchymal stem cell sheets.http://www.sciencedirect.com/science/article/pii/S2352320425000045Cell sheetThermoresponsive culture dishPolymer coating methodPeriodontitisPeriodontal ligament derived mesenchymal stem cell
spellingShingle Kazuki Morita
Masamichi Nakayama
Jiacheng Wang
Satoru Onizuka
Masahiro Hatasa
Yujin Ohsugi
Yosuke Tsuchiya
Hiromi Niimi
Anhao Liu
Hideaki Sakai
Teruo Okano
Takanori Iwata
Stable preparation of in vivo transplantable periodontal ligament-derived mesenchymal stem cell sheets in thermoresponsive culture dishes with tunable cell detachability
Regenerative Therapy
Cell sheet
Thermoresponsive culture dish
Polymer coating method
Periodontitis
Periodontal ligament derived mesenchymal stem cell
title Stable preparation of in vivo transplantable periodontal ligament-derived mesenchymal stem cell sheets in thermoresponsive culture dishes with tunable cell detachability
title_full Stable preparation of in vivo transplantable periodontal ligament-derived mesenchymal stem cell sheets in thermoresponsive culture dishes with tunable cell detachability
title_fullStr Stable preparation of in vivo transplantable periodontal ligament-derived mesenchymal stem cell sheets in thermoresponsive culture dishes with tunable cell detachability
title_full_unstemmed Stable preparation of in vivo transplantable periodontal ligament-derived mesenchymal stem cell sheets in thermoresponsive culture dishes with tunable cell detachability
title_short Stable preparation of in vivo transplantable periodontal ligament-derived mesenchymal stem cell sheets in thermoresponsive culture dishes with tunable cell detachability
title_sort stable preparation of in vivo transplantable periodontal ligament derived mesenchymal stem cell sheets in thermoresponsive culture dishes with tunable cell detachability
topic Cell sheet
Thermoresponsive culture dish
Polymer coating method
Periodontitis
Periodontal ligament derived mesenchymal stem cell
url http://www.sciencedirect.com/science/article/pii/S2352320425000045
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