| Project/Area Number |
15590907
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Neurology
|
|---|
| Research Institution | Keio University |
|---|
Principal Investigator |
KITA Yoshiko Keio University, Department of Medicine, 医学部, 講師 (00327496)
|
|---|
| Project Period (FY) |
2003 – 2004
|
| Project Status |
Completed (Fiscal Year 2004)
|
| Budget Amount *help |
¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 2004: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2003: ¥1,600,000 (Direct Cost: ¥1,600,000)
|
|---|
| Keywords | ES cells / Alzheimer's disease / Differentiation of ES cells / ALS / Kock-in / AD Disease model / APP / PS |
|---|
| Research Abstract |
Our final goal of this project is to create live neurons harboring Alzheimer' s disease(AD) abnormalities to study AD and to develop curative therapies. Basic strategy is that we constructed mouse embryonic stem(ES) cells, in which the AD-causative V642I mutation was introduced to the endogenous amyloid precursor protein(APP) gene, in combination with a protocol to efficiently differentiate ES cells into postmitotic neurons without using a cell sorter.
By this protocol, ES cells differentiated into more than 90% of the central type of adult postmitotic neurons.. The neurons derived from V642I-APP knock-in ES cells exhibited significantly increased Abeta 42 secretion.
Next, we created neurons derived from I213T-PS1 knock-in ES cell with the similar protocol. We analyzed gene expression of these mutation kock-in neurons using microarray analysis and validated with real time PCR. So far, expression of seven genes from neurons derived from V642I-APP and I213T-PS1 kock-in ES cells was increased compared with the one from wild type ES cells.
In addition, we are trying to create other neurodegenerative disease model neurons. We are working to prepare either G93R-SOD1 kock-in or transgenic ES cells to compare these neurodegenerative model neurons.. We are searching most effective differentiating protocol from the ES cells to motoneurons. Currently, we are co-culturing the ES cell derived neurons with sonic hedgehog producing cells to differentiate into motoneurons.
|
