2007 Fiscal Year Final Research Report Summary
Functional classification of bone marrow dtromal cells (BMSCs) using DNA microarray, and development ofculture methods which can maintain differentiation potential of BMSCs
| Project/Area Number |
17390414
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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 | Tokyo Medical and Dental University |
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Principal Investigator |
SHINOMIYA Kenichi Tokyo Medical and Dental University, Graduate SchoolOrthopaedic and Spinal Surgery, Professor (20111594)
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| Co-Investigator(Kenkyū-buntansha) |
AE Keisuke Tokyo Medical and Dental University, Graduate School, Orthopaedic and Spinal Surgery, Assistant Professor (20376726)
TAKEDA Shu Tokyo Medical and Dental University, Graduate School, Regenerative Therapeutics for Spine and Spinal Cord, Associate Professor (30376727)
SAOTOME Shinichi Tokyo Medical and Dental University, Graduate School, Regenerative Therapeutics for Spine and Spinal Cord, Associate Professor (20401391)
ENOMOTO Mitsuhiro Tokyo Medical and Dental University, Graduate School, Regenerative Therapeutics for Spine and Spinal Cord, Instructor (90451971)
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| Project Period (FY) |
2005 – 2007
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| Keywords | Bone marrow stromal cells / Bone regeneration / Differentiation capability / Dexamethasone / Cell seeding method / Plasma |
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| Research Abstract |
Human bone marrow stromal cells (hBMSCs) are an attractive source for bone tissue engineering applications because of their differentiation capability and their proliferation capability. However, despite the potential of BMSCs, they lose their differentiation potential after multiple doubling processes under standard culture conditions
1. We hypothesized that dexamethasone could augment the responsiveness of BMSCs to differentiation reagents, and continuous dexamethasone treatment would establish BMSCs with higher differentiation potentials. The results of our studies indicated that the dexamethasone-treated cells showed higher osteogenic capability and bone formation capability than the control group.
2. To develop efficient bone regeneration using cells and porous scaffolds, it is essential to introduce an adequate number of cells into porous scaffolds. To improve cell seeding efficiency, we modified the previously reported cell seeding techniques using low-pressure conditions and devi
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sed a simple device for this method. The procedures are as follows. In the first step, porous β-TCP blocks are placed into a glass chamber, and in the second step, the air in the chamber is removed by a syringe to create a vacuum. In the third step, the cell suspension is slowly infused into the chamber, maintaining the low-pressure condition, and the blocks are soaked in the suspension. In the final step, the valve is released so that the pressure in the chamber normalizes. Compared to spontaneous penetration (control), cell seeding efficiencies by this method were three fold higher, and the implants prepared by the method showed bone formation more than three folds of the control.
3. We also developed and tested a cell seeding system into porous implants using plasma, expecting that the fibrin network formed from fibrinogen would hold introduced cells within minutes and promote bone formation. BMSCs were suspended in plasma and introduced into porous scaffolds, and fibrin gel was formed within minutes. Then the implants were transplanted. The implants prepared using plasma showed significantly more bone formation than that by the implants using BMSCs suspended in culture medium. Less
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