| RRC ID |
62158
|
| Author |
Muramatsu M, Hanazono Y, Ogasawara Y, Okada T, Mizukami H, Kume A, Mizoguchi H, Ozawa K.
|
| Title |
Reversible integration of the dominant negative retinoid receptor gene for ex vivo expansion of hematopoietic stem/progenitor cells.
|
| Journal |
Biochem Biophys Res Commun
|
| Abstract |
Since hematopoietic stem cells (HSCs) differentiate readily ex vivo resulting in the loss of self-renewal and engraftment abilities, the transient block of differentiation is essential to maintain those abilities during their ex vivo expansion culture. To this end, we developed a method of reversible integration of the dominant negative retinoic acid receptor (DN-RAR) gene, a differentiation-blocking gene, into cells utilizing the Cre/loxP-dependent gene recombination system. The murine immature hematopoietic 32D cells differentiate into mature neutrophils upon G-CSF treatment. However, 32D cells transduced with a retroviral vector expressing the DN-RAR gene put between two loxP sites continued to proliferate without showing differentiation even in the presence of G-CSF. After the cells were fully amplified, the cells were transduced with the Cre recombinase gene. The cells then restored the ability to differentiate into mature neutrophils upon G-CSF treatment. PCR analysis showed that the DN-RAR gene was efficiently removed from the genome by introduction of the Cre gene. This system may eventually be applicable to the ex vivo expansion of HSCs.
|
| Volume |
285(4)
|
| Pages |
891-6
|
| Published |
2001-7-27
|
| DOI |
10.1006/bbrc.2001.5264
|
| PII |
S0006-291X(01)95264-7
|
| PMID |
11467834
|
| MeSH |
Animals
Cell Culture Techniques / methods
Cell Differentiation
Cell Division
Hematopoietic Stem Cells / cytology*
Integrases
Mice
Neutrophils / cytology*
Receptors, Retinoic Acid / genetics*
Recombination, Genetic
Transformation, Genetic
Viral Proteins
|
| IF |
2.985
|
| Resource |
| Human and Animal Cells |
10T1/2(RCB0247) |
