| Project/Area Number |
07670906
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Pediatrics
|
|---|
| Research Institution | Tokyo Women's Medical College |
|---|
Principal Investigator |
SAITO Kayoko Tokyo Women's Medical College, Dept.of Pediatrics, Associate Professor, 医学部, 助教授 (90138834)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
IKEYA Kiyoko Tokyo Women's Medical College, Dept.of Pediatrics, Assistant Professor, 医学部, 講師 (70151313)
近藤 恵里 東京女子医科大学, 医学部, 助手 (40281406)
|
|---|
| Project Period (FY) |
1995 – 1997
|
| Project Status |
Completed (Fiscal Year 1997)
|
| Budget Amount *help |
¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 1997: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1996: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1995: ¥1,300,000 (Direct Cost: ¥1,300,000)
|
|---|
| Keywords | progressive muscular dystrophy / Duchenne muscular dystrophy / Fukuyama muscular dystrophy / somatic mosaicism / gene therapy / cationic liposomes / cultured myoblasts / 体細胞モザイク / Duchenne型筋ジストロフィー / 転写産物 / 遺伝子産物 / Sequence / 骨格筋細胞培養 / アデノウイルスベクター |
|---|
| Research Abstract |
We have conducted the molecular genetic analysis and clinical application for genotype-phenotype correlation in progressive muscular dystrophies, especially Duchenne muscular dystrophy (DMD) and Fukuyama congenital muscular dystrophy (FCMD). For the purpose of obtaining complinentary primary genetic defects in the DMD gene by introducing copies of recombinant gene constructs into nuscle cells ex vivo, the potential use of cationic liposomes as physical gene delivery systems for cultured human dystrophic skeletal muscles was examined, from the basic scientific point of view. Various cationic liposome formulations, i.e.cellfectin, DMRIE-C,lipofectin, lipofectAMINE and lipofectAMINE PLUS,were examined for their use in plasmid DNA (beta-gal reporter gene) transfection. For transgene expression, muscle cultures were stained or were applied to detect beta-gal activities by the ELISA method. These in vitro studies indicated that lipofectAMINE PLUS showed high efficiency. In addition, myoblast non-clonal cultures were more efficient than clonal cultures. This is the first successful introduction of foreign genes into primary cultures of human dystrophic myoblasts by means of cationic liposomes.
|
