| Project/Area Number |
13671491
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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 | Chiba University |
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Principal Investigator |
WADA Yuichi Chiba University, University Hospital, Assistant, 医学部附属病院, 助手 (10282485)
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| Co-Investigator(Kenkyū-buntansha) |
TAKASE Kan Chiba University, Graduate School of Medicine, Assistant, 大学院・医学研究院, 助手 (40333489)
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| Project Period (FY) |
2001 – 2002
|
| Project Status |
Completed (Fiscal Year 2002)
|
| Budget Amount *help |
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2002: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2001: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | hyaluronan / gene transfer / chondrocyte implantation / ATDC5 / retrovirus vector / ヒアルロン酸合成酵素 |
|---|
| Research Abstract |
In the present study, plasmids containg HAS2 under the PMX-IRES-GFP retrovirus vector were transfected into undifferentiated ATDC5. As a control, cells transfected with the empty vector were used. ATDC5 cells differentiated in the presence of insulin and RT-PCR was performed to detect HAS2 m RNA expression and we evaluated visualization of pericellular coats which were composed of newly synthesized HA using by immunofluorescent staining and analyzed HA concentration in culture medium by high performance liquid chromatography. The signal of the cells transfected with HAS2 was visually much stronger than that of control cells. The HA concentration of the cells transfected with HAS2 in culture medium was much higher than that of control throughout 21 days. By transfection of HAS2 with retrovirus, we are able to expect the continuous synthesis of HA. Therefore this technique may be a great significance in autologous chondrocyte implantation.
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