| Project/Area Number |
18591431
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
General surgery
|
|---|
| Research Institution | Chiba University |
|---|
Principal Investigator |
MITSUHASHI Noboru Chiba University, Research Center for Frontier Medical Engineering, Associate Professor (80400985)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
MIYAZAKI Masaru Chiba University, Graduate School of Medicine, Professor (70166156)
KIMURA Fumio Chiba University, Graduate School of Medicine, Associate Professor (70334208)
SHIMIZU Hiroaki Chiba University, Graduate School of Medicine, Associate Professor (80272318)
YOSHIDOME Hiroyuki Chiba University, Graduate School of Medicine, Associate Professor (10312935)
OHTSUKA Masayuki Chiba University, Graduate School of Medicine, Assistant Professor (90334185)
|
|---|
| Project Period (FY) |
2006 – 2007
|
| Project Status |
Completed (Fiscal Year 2007)
|
| Budget Amount *help |
¥4,010,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥510,000)
Fiscal Year 2007: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Fiscal Year 2006: ¥1,800,000 (Direct Cost: ¥1,800,000)
|
|---|
| Keywords | Bone Marrow Stem Cell / VEGF / AFP / Homing |
|---|
| Research Abstract |
In this study, we attempted to establish a novel gene therapy with bone marrow mesenchymal stem cell. At first, we compared mRNA expression of chemokines, growth factors, and those receptors using quantitative RT-PCR between human hepatocellular carcinoma and adjacent tissues. We found difference of several genes expression. As a target gene for treatment, we selected alpha-fetoprotein (AFP) because AFP has properties of cell proliferation and anti-apoptosis. The expression of AFP in AFP transfected cell was confirmed by immunohistochemistry and enzyme linked immuno-sorbent assay (ELISA). AFP has confirmed as a stimulator of proliferation and an inhibitor of apoptosis of bone marrow MSC. As a target gene for homing, we selected VEGF/VEGFR-1 system because VEGF has known as a potent chemoattractant for bone marrow MSC. We also confirmed the expression of VEGFR-1 in mouse bone marrow MSC in protein level by flow cytometery. VEGF simulated migration of mouse bone marrow MSC in vitro. The double transfection of VEGFR-1 and AFP was performed using retrovirus vector. The double transfected cells showed stable expression of both VEGFR-1 and AFP assessed by flow cytometry and ELISA. Migration of double transfected cells to VEGF was confirmed using migration assay in vitro. Then, we performed subcutaneous injection of VEGF at back of mouse and intra venous injection of double transfected cells. Accumulation of double transfected cells at the back was confirmed by histological examination. Further, we confirmed migrated double transfected cells expressed AFP by histrological examination and ELISA. In conclusion, we established double transfected cells that could home to the lesion and produce the therapeutic molecule. This system may be a useful strategy for various kinds of diseases.
|
