Highly efficient transduction of repopulating bone marrow cells using rapidly concentrated polymer-complexed retrovirus.
Biochem Biophys Res Commun
; 330(3): 768-75, 2005 May 13.
Article
em En
| MEDLINE
| ID: mdl-15809063
ABSTRACT
Using the cationic polymer, Polybrene, and the anionic polymer, chondroitin sulfate C, we concentrated recombinant retrovirus pseudotyped with an ecotropic envelope, which is susceptible to inactivation by high-speed concentration methods. To evaluate gene marking, murine bone marrow was harvested from C3H mice, transduced with polymer-concentrated GFP virus, and transplanted into lethally irradiated recipients. Total gene marking in mice averaged 30-35% at 8 weeks post-transplant and transgene expression remained stable for over 16 weeks. Using the polymer concentration method, a second retroviral vector encoding the drug resistant variant of dihydrofolate reductase (L22Y-DHFR) was concentrated and tested. Approximately 40% of transduced murine bone marrow progenitor cells were protected against trimetrexate concentrations that completely eliminated the growth of non-modified cells. These results show that anionic and cationic polymers can be combined to rapidly concentrate viruses that are normally difficult to concentrate, and the concentrated virus efficiently transduces hematopoietic stem cells.
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Base de dados:
MEDLINE
Assunto principal:
Polímeros
/
Retroviridae
/
Transdução Genética
/
Células da Medula Óssea
Idioma:
En
Ano de publicação:
2005
Tipo de documento:
Article