2004 Fiscal Year Final Research Report Summary
Cartilage differentiation and repair from mesenchymal stem cells using gene transfer of Hyaluronan synthase 2
| Project/Area Number |
14370451
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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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 | Chiba University |
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Principal Investigator |
MORIYA Hideshige CHIBA University, Graduate School of Medicine, Department of Orthopaedic Surgery, Professor, 大学院・医学研究院, 教授 (30092109)
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| Co-Investigator(Kenkyū-buntansha) |
WADA Yuichi Teikyo Uneversity School of Medicine Department of Orthopaedic Surgery, Ichihara Hospital, Professor, 医学部附属病院, 教授 (10282485)
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| Project Period (FY) |
2002 – 2004
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| Keywords | Cartilage / ATDC5 / Hyaluronan / Hyaluronan synthase 2 / Retrovirus vector / Gene transfer |
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| Research Abstract |
In 2002,we were able to make out the model of all chondrogenic differentiation by culture of the mouse chondrogenic cell line ATDC5, on the basis of chondrogenic potential in the presence of insulin.
In 2003,we succeeded in gene transfer of Hyaluronan synthase 2 (HAS2) to ATDC5 using retrovirus vector. The transfectants by this technique were able to expect continuous synthesis of HA throughout the chondrogenic differentiaion,and as a result, it became to produce of extracellular matrixs of cartilage. We performed isolation and culture of mesenchymal stem cell to have the capacity of chondrogenic differentiation from the myeloid tissue of pure rabbits iliac bone, and performed retrovirus-mediated gene transfer method of HAS2 to ATDC5 and finally were able to culture of these cells into collagen gels.
In 2004,we performed the transplantation of above-mentioned stem cells into collagen gels in the femoral component of the rabbit knee and patched on periosteal grafts which were obtained from the proximal of tibia of rabbits, as a fact,we were able to make out the models of the cartilage repair. In this cartilage repair models, at 4 weeks after grafting, HAS2 transfected groups (the group of retrovirus vector and the group of gene gun) showed much higher levels of HA synthesis compared with HA untransfected cells group using by irnmunofluorescent staining. And so, at 12 weeks after surgery, RT-PCR analysis revealed higher expression of type 2 collagen in the HAS2 groups and surface regularity and total histological score in the HAS2 groups were significantly better than those in control.
In these effects, HAS2 gene transfer had the possibility of the hyaline cartilage repair.
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