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Gene Marking in Adeno-Associated Virus Vector Infected Periosteum Derived Cells for Cartilage Repair
NAOMI KOBAYASHI, TOMIHISA KOSHINO, MASAAKI UESUGI, NAOKI YOKOO, KE-QIN XIN, KENJI OKUDA, HIROAKI MIZUKAMI, KEIYA OZAWA, and TOMOYUKI SAITO
ABSTRACT.
Methods. Periosteum was obtained from the tibia of 6-week-old rabbits and enzymatically digested. The isolated periosteum derived cells were cultured and the subconfluence cells were infected with a recombinant AAV expressing the LacZ gene (AAV-LacZ). Collagen gel containing the LacZ transferred, periosteum derived cells was transplanted into a full thickness articular cartilage defect in 10 rabbits. Results. Infected cells still growing on the plate continued to express LacZ at least 12 weeks after AAV infection, with the highest percentage of LacZ positive cells reaching 74.4%. The LacZ positive cells were recognized at the transplant sites in 8 out of 10 knees. Conclusion. Gene expression in periosteum derived cells was sustained in vitro for at least 12 weeks using the AAV vector, and for 2 weeks ex vivo after transplantation into a cartilage defect. (J Rheumatol 2002;29:2176-80) Key Indexing Terms:
ADENO-ASSOCIATED VIRUS
From the Departments of Orthopaedic Surgery and Bacteriology, Yokohama City University School of Medicine, Yokohama; and Division of Genetic Therapeutics, Center for Molecular Medicine, Jichi Medical School, Yokohama, Japan. Supported by a Grant-in-Aid from the Ministry of Education, Science, Sports, and Culture of Japan. N. Kobayashi, MD; T. Koshino, MD, PhD; M. Uesugi, MD, PhD; N. Yokoo, MD, Department of Orthopaedic Surgery; K. Xin, MD, PhD; K. Okuda, MD, PhD, Department of Bacteriology, Yokohama City University School of Medicine; H. Mizukami, MD, PhD; K. Ozawa, MD, PhD, Division of Genetic Therapeutics, Center for Molecular Medicine, Jichi Medical School; T. Saito, MD, PhD, Department of Orthopaedic Surgery, Yokohama City University School of Medicine. Address reprint requests to Dr. N. Kobayashi, Department of Orthopaedic Surgery, Yokohama City University School of Medicine, 3-9 Fukuura, Kanazawa-ku, Yokohama 236-0004, Japan. E-mail: naomik@ea.mbn.or.jp Submitted August 13, 2001; revision accepted April 24, 2002. |