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Blood, 15 January 2002, Vol. 99, No. 2, pp. 709-712
BRIEF REPORT
Lentiviral gene transfer into peripheral blood-derived
CD34+ NOD/SCID-repopulating cells
Michaela Scherr,
Karin Battmer,
Ulrike Blömer,
Bernd Schiedlmeier,
Arnold Ganser,
Manuel Grez, and
Matthias Eder
From the Hannover Medical School, Department Hematology
and Oncology, Hannover, Germany; Heinrich-Pette Institute, Hamburg,
Germany; and Institute for Biomedical Research, Georg-Speyer-Haus,
Frankfurt, Germany.
This study reports a lentiviral gene transfer protocol for
efficient transduction of adult human peripheral blood
(PB)-derived CD34+ NOD/SCID-repopulating cells (SRCs)
using vesicular stomatitis virus-G protein (VSV-G)-pseudotyped
lentiviruses encoding for enhanced green fluorescence protein
(eGFP). Lentiviral stocks were concentrated by anion exchange
chromatography, and transduction was performed under serum-free
conditions at a multiplicity of infection (MOI) between 3 and 50. Similar transduction efficiencies were achieved in the presence and
absence of cytokines. Transduction of PB-derived CD34+
cells at a MOI of 3 resulted in gene transfer efficiencies into SRCs of
9.2% and 12.0% in the absence and presence of cytokines, respectively. Using improved lentiviral vectors, transduction frequency
varied between 42.0% (MOI 10) and 36.0% (MOI 50) with multilineage
transgene expression within SRC-derived myeloid and lymphoid cells. The
protocol described can be adapted for clinical application of
lentiviral gene transfer into PB-derived CD34+ cells from
adult patients.
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