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2006 | 53 | 4 | 815-823
Article title

Optimization of a retroviral vector for transduction of human CD34 positive cells

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EN
Abstracts
EN
Human stem and progenitor cells have recently become objects of intensive studies as an important target for gene therapy and regenerative medicine. Retroviral vectors are among the most effective tools for genetic modification of these cells. However, their transduction efficiency strongly depends on the choice of the ex vivo transduction system. The aim of this study was to elaborate a system for retroviral vector transduction of human CD34 positive cells isolated from cord blood. The retroviral vector pMINV EGFP was chosen for transduction of two human erythroblastoid cell lines: KG-1a (CD34 positive) and K562 (CD34 negative). For vector construction, three promoters and two retroviral vector packaging cell lines were used. To optimize the physicochemical conditions of the transduction process, different temperatures of supernatant harvesting, the influence of centrifugation and the presence of transduction enhancing agents were tested. The conditions elaborated with KG-1a cells were further applied for transduction of CD34 positive cells isolated from cord blood. The optimal efficiency of transduction of CD34 positive cells with pMINV EGFP retroviral vector (26% of EGFP positive cells), was obtained using infective vector with LTR retroviral promoter, produced by TE FLY GA MINV EGFP packaging cell line. The transduction was performed in the presence of serum, at 37°C, with co-centrifugation of cells with viral supernatants and the use of transduction enhancing agents. This study confirmed that for gene transfer into CD34 positive cells, the detailed optimization of each element of the transduction process is of great importance.
Publisher

Year
Volume
53
Issue
4
Pages
815-823
Physical description
Dates
published
2006
received
2006-07-03
revised
2006-10-31
accepted
2006-12-07
(unknown)
2006-12-11
Contributors
author
  • Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wrocław, Poland
  • Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wrocław, Poland
  • Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wrocław, Poland
  • Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wrocław, Poland
  • Chair of Obstetrics and Gynecology of Wrocław Medical University, Wrocław, Poland
  • Department of Cancer Immunology, Chair of Oncology, University of Medical Sciences at Great Poland Cancer Center, Poznań, Poland
  • Department of Cancer Immunology, Chair of Oncology, University of Medical Sciences at Great Poland Cancer Center, Poznań, Poland
author
  • Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wrocław, Poland
References
  • Byk T, Haddada H, Vainchenker W, Louache F (1998) Lipofectamine and related cationic lipids strongly improve adenoviral infection efficiency of primitive human hematopoietic cells. Hum Gene Ther 9: 2493-2502.
  • Chuck AS, Clarke MF, Palsson BO (1996) Retroviral infection is limited by brownian motion. Hum Gene Ther 7: 1527-1534.
  • Coffin JM, Hughes SH, Varmus HE (eds) (1997) Retroviruses. Cold Spring Harbor Laboratory Press: Cold Spring Harbor, New York.
  • Darling D, Huges C, Galea-Lauri J, Gäken J, Trayner ID, Kuiper M, Farzaneh F (2000) Low-speed centrifugation of retroviral vectors absorbed to a particulate substrate: a higly effective means of enhancing retroviral titre. Gene Ther 7: 914-923.
  • Dropulic B (2005) Genetic modification of hematopoietic cells using retroviral and lentiviral vectors: safety considerations for vector design and delivery into target cells. Curr Hematol Rep 4: 300-304.
  • Forestell SP, Böhnlein E, Rigg R.J (1995) Retroviral end-point titer is not predictive of gene tranfer efficiency implications for vector production. Gene Ther 2: 723-730.
  • Grabarczyk P, Wysocki PJ, Gryska K, Mackiewicz A (2002) Expression of PiT1 and PiT2 receptors and transduction efficiency of tumor cels. Acta Biochim Polon 49: 333-339.
  • Grignani F, Kinsella T, Mencarelli A, Valtieri M, Riganelli D, Grignani F, Lanfrancone L, Peschle C, Nolan GP, Pelicci PG (1998) High - efficiency gene transfer and selection of human haematopoietic progenitor cells with a hybrid EBV/retroviral vector expressing the green fluorescence protein. Cancer Res 58: 14-19.
  • Hawley RG, Lieu FH, Fong AZ, Hawley TS (1994) Versatile retroviral vectors for potential use in gene therapy. Gene Ther 1: 136-138.
  • Hong Y, Lee K, Yu SS, Kim S, Kim JG, Shin HY, Kim S (2004) Factors affecting retrovirus-mediated gene transfer to human CD34+cells. J Gene Med 6: 724-733.
  • Kaiser J (2005) American Society of Gene Therapy meeting. Retroviral vectors: a double-edged sword. Science 308: 1735-1736.
  • Kaptein LC, Greijer AE, Valerio D, van Beusechem VW (1997) Optimized conditions for the production of recombinant amphotropic retroviral vector preparations. Gene Ther 4: 172-176.
  • Kim SH, Kim S, Robbins PD (2000) Retroviral vectors. Adv Virus Res 55: 545-563.
  • Kotani H, Newton III PB, Zhang S, Chiang YL, Otto E, Weaver L, Blaese M, Anderson WF, McGarrity GJ (1994) Improved methods for retroviral vector transduction and production for gene therapy. Hum Gene Ther 5: 19-28.
  • Lee CGL, Pastan I, Gottesman MM (1998) Retroviral transfer of human MDR1 gene into human T lymphocytes. Methods Enzymol 292: 557-572.
  • Liu H, Hung Y, Wissink SD, Verfaillie CM (2000) Improved retroviral transduction of haematopoietic progenitors by combining methods to enhance virus-cell interaction. Leukemia 14: 307-311.
  • MacNeil EC, Hanenberg H, Pollok KE, van der Loo JC, Bierhuizen MF, Wagemaker G, Williams DA (1999) Simultaneous infection with retroviruses pseudotyped with different envelope proteins bypasses viral receptor interference associated with colocalization of gp70 and target cells on fibronectin CH-296. J Virol 73: 3960-3967.
  • Reeves L, Cornetta K (2000) Clinical retroviral vector production by step filtration using clinically approved filters improves titers. Gene Ther 7: 1993-1998.
  • Salmon P, Kindler V, Ducrey O, Chapuis B, Zubler RH and Trono D (2000) High-level transgene expression in human hematopoietic progenitors and differentiated blood lineages after transduction with improved lentiviral vectors. Blood 96: 3392-3398.
  • Schiedlmeier B, Kühlcke K, Eckert HG, Baum C, Zeller WJ, Fruehauf S (2000) Quantitative assessment of retroviral transfer of the human multidrug resistance 1 gene to human mobilized peripheral blood progenitor cells engrafted in nonobese diabetic/severe combined immunodeficient mice. Blood 95: 1237-1248.
  • Schmidt M, Hacein-Bey-Abina S, Wissler, M, Carlier F, Lim A, Prinz C, Glimm H, Andre-Schmutz I, Hue C, Garrigue A, Le Deist F, Lagresle C, Fischer A, Cavazzana-Calvo M, von Kalle C (2005) Clonal evidence for the transduction of CD34+cells with lymphomyeloid differentiation potential and self-renewal capacity in the SCID-X1 gene therapy trial. Blood 105: 2699-2706.
  • Thomsen S, Vogt B,von Laer D, Heberlein C, Rein A, Ostertag W, Stocking C (1998) Lack of functional PiT-1 and PiT-2 expression on hematopoietic stem cell lines. Acta Haematol 99: 148-155.
  • Woods NB, Mikkola H, Nilsson E, Olsson K, Trono D, Karlsson S (2001) Lentiviral-mediated gene transfer into haematopoietic stem cells. J Intern Med 249: 339-343.
Document Type
Publication order reference
Identifiers
YADDA identifier
bwmeta1.element.bwnjournal-article-abpv53p815kz
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