Highly efficient retroviral gene transfer based on centrifugation-mediated vector preloading of tissue culture vessels.
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Highly efficient retroviral gene transfer based on centrifugation-mediated vector preloading of tissue culture vessels. / Kühlcke, Klaus; Fehse, Boris; Schilz, Andrea; Loges, Sonja; Lindemann, Carsten; Ayuketang Ayuk, Francis; Lehmann, Friederike; Stute, Norbert; Fauser, Axel A; Zander, Axel R; Eckert, Hans-Georg.
in: MOL THER, Jahrgang 5, Nr. 4, 4, 2002, S. 473-478.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
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TY - JOUR
T1 - Highly efficient retroviral gene transfer based on centrifugation-mediated vector preloading of tissue culture vessels.
AU - Kühlcke, Klaus
AU - Fehse, Boris
AU - Schilz, Andrea
AU - Loges, Sonja
AU - Lindemann, Carsten
AU - Ayuketang Ayuk, Francis
AU - Lehmann, Friederike
AU - Stute, Norbert
AU - Fauser, Axel A
AU - Zander, Axel R
AU - Eckert, Hans-Georg
PY - 2002
Y1 - 2002
N2 - Efficient retroviral gene transfer into primary cells is a prerequisite for various gene therapeutic strategies. We have developed a transduction protocol based on the preloading of tissue culture vessels with retroviral particles by low-speed (1000g) centrifugation. We show that vector-preloaded tissue culture vessels allow highly efficient gene transfer into various target cells. We obtained transduction rates of up to 85% for primary T lymphocytes after just a single round of transduction. Under clinically relevant conditions using a vector developed for suicide gene therapy and produced under good manufacturing practice (GMP) conditions, the described method allowed generation of large numbers (>2x10(9)) of gene-modified T cells. The preloading concept ensures transduction of target cells in their optimal growth medium regardless of the medium used for vector production. This facilitated highly efficient gene transfer into quite different target cells such as CD34(+) and AC133(+) bone marrow progenitor as well as mesenchymal stem cells. The presented method combines high gene-transfer rates with a great potential for standardization in accordance with GMP guidelines and is consequently well suited for both research and clinical applications. (c)2002 Elsevier Science (USA).
AB - Efficient retroviral gene transfer into primary cells is a prerequisite for various gene therapeutic strategies. We have developed a transduction protocol based on the preloading of tissue culture vessels with retroviral particles by low-speed (1000g) centrifugation. We show that vector-preloaded tissue culture vessels allow highly efficient gene transfer into various target cells. We obtained transduction rates of up to 85% for primary T lymphocytes after just a single round of transduction. Under clinically relevant conditions using a vector developed for suicide gene therapy and produced under good manufacturing practice (GMP) conditions, the described method allowed generation of large numbers (>2x10(9)) of gene-modified T cells. The preloading concept ensures transduction of target cells in their optimal growth medium regardless of the medium used for vector production. This facilitated highly efficient gene transfer into quite different target cells such as CD34(+) and AC133(+) bone marrow progenitor as well as mesenchymal stem cells. The presented method combines high gene-transfer rates with a great potential for standardization in accordance with GMP guidelines and is consequently well suited for both research and clinical applications. (c)2002 Elsevier Science (USA).
M3 - SCORING: Zeitschriftenaufsatz
VL - 5
SP - 473
EP - 478
JO - MOL THER
JF - MOL THER
SN - 1525-0016
IS - 4
M1 - 4
ER -
