2001 Fiscal Year Final Research Report Summary
Gene transfer to periosteum-derived cells for cartilage repair
| Project/Area Number |
12671432
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Orthopaedic surgery
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| Research Institution | Department of Orthpaedic Surgery, Yokohama city university |
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Principal Investigator |
KAWAI Kosei Yokohama city university Assistant, 医学部附属病院, 助手 (20305452)
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| Co-Investigator(Kenkyū-buntansha) |
XIN Ke-qin Yokohama city university Assistant, 医学部, 助手 (40301452)
OZAWA Keiya Jichi Medical School, 医学部, 教授 (30137707)
SAITO Tomoyuki Yokohama city university Assistant Proffesor, 医学部, 助教授 (30170517)
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| Project Period (FY) |
2000 – 2001
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| Keywords | periosteal cell / cartilage repair / AAV vector / TGF-β / LacZ |
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| Research Abstract |
<INTRODUCTION>
Recently, periosteum-derived cells are known to have chondrogenic potential and used as a substitute for chondrocytes for cartilage defect. Adeno-associate virus(AAV) vector is well recogniged as exquisite vector that infect easily to many kinds of cells and have no virulency. We investigated the ability of gene transfer to periosteal cells using AAV vector including LacZ gene and the cytotoxicity of AAV vector to periosteal cell.
<MATERIAL ＆ METHODS>
Periosteum were harvestsd from tibiae of 6 week-old rabbits. Periosteum derived cells were isolated by digestion with 0.25 % trypsin EDTA and 0.25 % collagenase. When cells reached subconfluence, AAV including LacZ gene was infected to cells. One week after infection, those cells were collected and cultured at high densities(4.0×10^<16> cells/ml) in 0.25 %collagen gel for 1 week. Collagen gells with the cells were transplanted into full thickness defect of articular cartilage(diameter 5mm, depth 3mm) made at the knee of 8 rabbits. Two to six weeks after the grafting, the surface of implanted site was observed, and the distal end of the femur was obtained and stained with X-gal staining to quantify Lac Z expression.
<RESULTS>
The expression of Lac Z was confirmed at 2, 4, 6 weeks after the grafting at the defect of articular cartilage in all of 8 rabbits. There was no side effects to transplanted cells.
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