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Titolo:
Safe sorting of GFP-transduced live cells for subsequent culture using a modified FACS vantage
Autore:
Sorensen, TU; Gram, GJ; Nielsen, SD; Hansen, JES;
Indirizzi:
HS Hvidovre Hosp, Dept Infect Dis 144, DK-2650 Hvidovre, Denmark HS Hvidovre Hosp Hvidovre Denmark DK-2650 144, DK-2650 Hvidovre, Denmark
Titolo Testata:
CYTOMETRY
fascicolo: 4, volume: 37, anno: 1999,
pagine: 284 - 290
SICI:
0196-4763(199912)37:4<284:SSOGLC>2.0.ZU;2-3
Fonte:
ISI
Lingua:
ENG
Soggetto:
GREEN-FLUORESCENT PROTEIN; RETROVIRAL VECTOR; GENE-EXPRESSION; METHYLATION;
Keywords:
flow cytometry; cell sorting; safety; retroviral transduction; green fluorescent protein;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
15
Recensione:
Indirizzi per estratti:
Indirizzo: Gram, GJ HS Hvidovre Hosp, Dept Infect Dis 144, Kettegard Alle 30, DK-2650Hvidovre, Denmark HS Hvidovre Hosp Kettegard Alle 30 Hvidovre Denmark DK-2650 mark
Citazione:
T.U. Sorensen et al., "Safe sorting of GFP-transduced live cells for subsequent culture using a modified FACS vantage", CYTOMETRY, 37(4), 1999, pp. 284-290

Abstract

Background: A stream-in-air cell sorter enables rapid sorting to a high purity, but it is not well suited for sorting of infectious material due to the risk of airborne spread to the surroundings. Methods: A FACS Vantage cell sorter was modified for safe use with potentially HIV infected cells. Safety tests with bacteriophages were performed toevaluate the potential spread of biologically active material during cell sorting. Cells transduced with a retroviral vector carrying the gene for GFP were sorted on the basis of their GFP fluorescence, and GFP expression was followed during subsequent culture. Results: The bacteriophage sorting showed that re biologically active material was confined to the sorting chamber. A failure mode simulating a nozzle blockage resulted in detectable droplets inside the sorting chamber, but no droplets could be detected when an additional air suction from the sorting chamber had been put on. The GFP transduced cells were sorted to 99% purity. Cells not expressing GFP at the time of sorting did not turn on the gene during subsequent culture. Unsorted cells and cells sorted to be positive for GFP showed a decrease in the fraction of GFP positive cells during culture. Conclusions: Sorting of live infected cells can be performed safely and with no deleterious effects on vector expression using the modified FAGS Vantage instrument. Cytometry 37. 284-290, 1999. (C) 1999 Wiley-Liss, Inc.

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Documento generato il 27/11/20 alle ore 06:26:10