2021 Fiscal Year Final Research Report

Osteochondral regeneration by autologous transplantation of abundant stem cells cultured high-densely

Research Project

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Project/Area Number	19H03129
Research Category	Grant-in-Aid for Scientific Research (B)
Allocation Type	Single-year Grants
Section	一般
Review Section	Basic Section 42020:Veterinary medical science-related
Research Institution	Kagoshima University
Principal Investigator	MISUMI KAZUHIRO 鹿児島大学, 農水産獣医学域獣医学系, 教授 (10291551)
Co-Investigator(Kenkyū-buntansha)	藏元智英鹿児島大学, 農水産獣医学域獣医学系, 助教 (80813154) 畠添孝鹿児島大学, 農水産獣医学域獣医学系, 准教授 (90776874)
Project Period (FY)	2019-04-01 – 2022-03-31
Keywords	幹細胞 / 軟骨 / 骨 / 関節症 / 再生医療
Outline of Final Research Achievements	The aims of this study were to establish a method to obtain a large amount (1 billion cells) of equine synovium derived stem cells (SMMSC) with the minimum passage (passage 1: P1), and to carry out the clinical trial with autologous implantation of SMMSC. Passage 0-SMMSC (P0-SMMSC) were isolated and cultured from nuclear cells of synovium that was collected during surgery in equine intra-articular fracture cases, by using a automatic cell culture device containing a non-woven fabric of polyethylene terephthalate filaments. An average over 100 million P0-SMMSC were isolated by culturing for 10 to 13 days, and then one passage culture successfully made to expand the P0-SMMSCs 13 to 18 times (1.3-1.8 billion P1-SMMSC). In a clinical trial of autologous implantation, more than 100 million P0-SMMSCs that were obtained in 11-14 days with 10% autologous serum- added medium were intraarticularly administrated, no clinical side-effects were presented not only locally but also systemically.
Free Research Field	臨床獣医学
Academic Significance and Societal Importance of the Research Achievements	継代培養を重ねることによる体性幹細胞としての機能劣化に伴って、滑膜由来幹細胞（SMMSC）に特徴的な軟骨分化能は消退していく。SMMSCによる治療の有効性を高めるために、継代数を最小限に止めて幹細胞としての品質を保持した細胞を十分数得る技術が必要であった。本研究では、細胞が接着する表面積が大きいポリエチレンタフレートフィラメント不織布を用いた馬のSMMSCの大量培養に成功した。これにより関節鏡手術時に採取した滑膜組織と自己血清を用いて分離培養した１億個の自己由来初代SMMSCを術後２週間で関節に自家移植する治療が可能となった。関節内骨折症例で移植後の安全性を確認し、実用化に向け大きく進展した。