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Titolo:
High-efficiency transduction and long-term gene expression with a murine stem cell retroviral vector encoding the green fluorescent protein in human marrow stromal cells
Autore:
Marx, JC; Allay, JA; Persons, DA; Nooner, SA; Hargrove, PW; Kelly, PF; Vanin, EF; Horwitz, EM;
Indirizzi:
St Jude Childrens Res Hosp, Div Expt Hematol, Memphis, TN 38105 USA St Jude Childrens Res Hosp Memphis TN USA 38105 ol, Memphis, TN 38105 USA StNJude Childrens Res Hosp, Transplantat & Gene Therapy Program, Memphis, T St Jude Childrens Res Hosp Memphis TN USA 38105 erapy Program, Memphis, T
Titolo Testata:
HUMAN GENE THERAPY
fascicolo: 7, volume: 10, anno: 1999,
pagine: 1163 - 1173
SICI:
1043-0342(19990501)10:7<1163:HTALGE>2.0.ZU;2-N
Fonte:
ISI
Lingua:
ENG
Soggetto:
APE LEUKEMIA-VIRUS; HUMAN FACTOR-IX; IN-VIVO; HEMATOPOIETIC-CELLS; IMMUNODEFICIENT MICE; PHOSPHATE-DEPLETION; PERIPHERAL-BLOOD; HEMOPHILIA-A; BONE; PROGENITORS;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
48
Recensione:
Indirizzi per estratti:
Indirizzo: Horwitz, EM St Jude Childrens Res Hosp, Div Expt Hematol, 332 N Lauderdale,Room D-4026, St Jude Childrens Res Hosp 332 N Lauderdale,Room D-4026 Memphis TN USA 38105
Citazione:
J.C. Marx et al., "High-efficiency transduction and long-term gene expression with a murine stem cell retroviral vector encoding the green fluorescent protein in human marrow stromal cells", HUM GENE TH, 10(7), 1999, pp. 1163-1173

Abstract

Bone marrow stromal cells (MSCs) are unique mesenchymal cells that have been utilized as vehicles for the delivery of therapeutic proteins in gene therapy protocols. However, there are several unresolved issues regarding their potential therapeutic applications, These include low transduction efficiency, attenuation of transgene expression, and the technical problems associated with drug-based selection markers, To address these issues, we have developed a transduction protocol that yields high-level gene transfer intohuman MSCs, employing a murine stem cell virus-based bicistronic vector containing the green fluorescent protein (GFP) gene as a selectable marker, Transduction of MSCs plated at low density for 6 hr per day for 3 days with high-titer viral supernatant resulted in a gene transfer efficiency of 80 +/- 6% (n = 10) as measured by GFP fluorescence. Neither centrifugation nor phosphate depletion increased transduction efficiency, Assessment of amphotropic receptor (Pit-2) expression by RT-PCR demonstrated that all MSCs expressing the receptor were successfully transduced. Cell cycle distribution profiles measured by propidium iodide staining showed no correlation with the susceptibility of MSCs to transduction by the retroviral vector, Human MSCs sequentially transduced with an adenoviral vector encoding the ecotropicreceptor and ecotropic retroviral vector encoding GFP demonstrated that all MSCs are susceptible to retroviral transduction, We further showed that both genes of bicistronic vector are expressed for at least 6 months in vitro and that transgene expression did not affect the growth or osteogenic differentiation potential of MSCs. Future studies will be directed toward the development of gene therapy protocols employing this strategy.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 24/11/20 alle ore 14:25:44