| Project/Area Number |
14580774
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Neuroscience in general
|
|---|
| Research Institution | KEIO UNIVERSITY |
|---|
Principal Investigator |
NAMIKI Jun Keio University School of Medicine, Department of Emergency Medicine, Assistant Professor, 医学部, 専任講師 (20189195)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
SHIMAZAKI Takuya Keio University School of Medicine, Department of Physiology, Instructor, 医学部, 助手 (00324749)
OKANO Hideyuki Keio University School of Medicine, Department of Physiology, Professor, 医学部, 教授 (60160694)
|
|---|
| Project Period (FY) |
2002 – 2003
|
| Project Status |
Completed (Fiscal Year 2003)
|
| Budget Amount *help |
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2003: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 2002: ¥1,900,000 (Direct Cost: ¥1,900,000)
|
|---|
| Keywords | Neural Stem Cells / CNS Regeneration / Marrow Stromal Cells / Nestin / GFP / Cell Sorter / nestin |
|---|
| Research Abstract |
Although Nestin protein is known as a marker for the neural stem cell (NSC), it is not specific to NSC because it is expressed in cells other than NSC, for example, the vascular endothelial cell. The second intronic enhancer of the nestin gene regulates gene expression in NSC selectively. Therefore, NSC characteristics of cells obtained from the E-nestin: EGFP transgenic mouse in which expression of the second intronic enhancer of the nestin gene is visualized by the enhanced green fluorescent protein (EGFP) correlate to the fluorescent intensity of the GFP. We induced NSC-selective nestin gene expression in the marrow stromal cell (MSC) using fluorescence of GFP as an index. Furthermore, we isolated and enriched nestin-EGFP positive MSC by a cell sorter.
The mesodermal Nestin protein was expressed in bone marrow-derived cells. The bone marrow of the E/nestin: EGFP transgenic mouse enabled to visualize NSC-selective nestin gene expression. Noggin, an antagonist to BMP, could induce NSC-selective nestin gene expression in MSC. NSC-selective nestin gene-expressing MSCs were isolated and enriched by a cell sorter. These cells did not proliferate in the neurosphere assay of NSC nor express differentiation marker proteins of CNS. Phenotypes of cell surface antigens of NSC-selective nestin gene-expressing MSCs were CD45-, cKit-, Scal^<low/+>, CD31^-, CD44^-, and CD49e^-. These phenotypes were similar to those of multipotent adult progenitor cell (MAPC).
|
