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Selective immunoaffinity-based enrichment of CD34+ cells transduced with retroviral vectors containing an intracytoplasmatically truncated version of the human low-affinity nerve growth factor receptor (deltaLNGFR) gene

Standard

Selective immunoaffinity-based enrichment of CD34+ cells transduced with retroviral vectors containing an intracytoplasmatically truncated version of the human low-affinity nerve growth factor receptor (deltaLNGFR) gene. / Fehse, B; Uhde, A; Fehse, Natalja; Eckert, H G; Clausen, Johannes; Rüger, R; Koch, S; Ostertag, W; Zander, A R; Stockschläder, M.

In: HUM GENE THER, Vol. 8, No. 15, 10.10.1997, p. 1815-1824.

Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review

Harvard

Fehse, B, Uhde, A, Fehse, N, Eckert, HG, Clausen, J, Rüger, R, Koch, S, Ostertag, W, Zander, AR & Stockschläder, M 1997, 'Selective immunoaffinity-based enrichment of CD34+ cells transduced with retroviral vectors containing an intracytoplasmatically truncated version of the human low-affinity nerve growth factor receptor (deltaLNGFR) gene', HUM GENE THER, vol. 8, no. 15, pp. 1815-1824. https://doi.org/10.1089/hum.1997.8.15-1815

APA

Fehse, B., Uhde, A., Fehse, N., Eckert, H. G., Clausen, J., Rüger, R., Koch, S., Ostertag, W., Zander, A. R., & Stockschläder, M. (1997). Selective immunoaffinity-based enrichment of CD34+ cells transduced with retroviral vectors containing an intracytoplasmatically truncated version of the human low-affinity nerve growth factor receptor (deltaLNGFR) gene. HUM GENE THER, 8(15), 1815-1824. https://doi.org/10.1089/hum.1997.8.15-1815

Vancouver

Fehse B, Uhde A, Fehse N, Eckert HG, Clausen J, Rüger R et al. Selective immunoaffinity-based enrichment of CD34+ cells transduced with retroviral vectors containing an intracytoplasmatically truncated version of the human low-affinity nerve growth factor receptor (deltaLNGFR) gene. HUM GENE THER. 1997 Oct 10;8(15):1815-1824. https://doi.org/10.1089/hum.1997.8.15-1815

Bibtex

@article{25960db7c78648bdaff7972348fe4872,
title = "Selective immunoaffinity-based enrichment of CD34+ cells transduced with retroviral vectors containing an intracytoplasmatically truncated version of the human low-affinity nerve growth factor receptor (deltaLNGFR) gene",
abstract = "Human hematopoietic stem cells remain one of the most promising target cells for gene therapeutic approaches to treat malignant and nonmalignant diseases. To rapidly characterize transduced cells and to isolate these from residual nontransduced, but biologically equivalent, cells, we have used a Moloney murine leukemia virus (Mo-MuLV)-based retroviral vector containing the intracytoplasmatically truncated human low-affinity nerve growth factor receptor (deltaLNGFR) cDNA as a marker gene. Supernatant transduction of CD34+ cells (mean purity 97%) in fibronectin-coated tissue culture flasks resulted in 5.5-45% (mean 26%) transduced cells expressing deltaLNGFR (LNGFR+ cells). After transduction, more than 65% of the transduced cells remained CD34+. Compared with control (mock- and nontransduced) CD34+ cells, transduction did not decrease the cloning efficiency of CD34+ cells. Immunomagnetic selection of the transduced cells with a monoclonal anti-LNGFR antibody resulted in >90% LNGFR+ cells. Further phenotypic characterization of these highly enriched LNGFR+ cells indicated that the majority co-expressed the CD34 and CD38 antigens. These results show that transduced cells expressing an ectopic cell-surface protein can be rapidly and conveniently quantitated and characterized by fluorescence-activated cell sorting (FACS) analysis and fast and efficiently enriched by immunoadhesion using magnetic beads. The use of cell-surface reporters should facilitate optimization of methods of gene transfer into more primitive hematopoietic progenitors.",
keywords = "Animals, Antigens, CD34, Cloning, Molecular, Fibronectins/pharmacology, Gene Transfer Techniques, Genetic Vectors, Hematopoietic Stem Cells/immunology, Humans, Mice, Moloney murine leukemia virus/genetics, Receptors, Nerve Growth Factor/biosynthesis, Time Factors, Transformation, Genetic",
author = "B Fehse and A Uhde and Natalja Fehse and Eckert, {H G} and Johannes Clausen and R R{\"u}ger and S Koch and W Ostertag and Zander, {A R} and M Stockschl{\"a}der",
year = "1997",
month = oct,
day = "10",
doi = "10.1089/hum.1997.8.15-1815",
language = "English",
volume = "8",
pages = "1815--1824",
journal = "HUM GENE THER",
issn = "1043-0342",
publisher = "Mary Ann Liebert Inc.",
number = "15",

}

RIS

TY - JOUR

T1 - Selective immunoaffinity-based enrichment of CD34+ cells transduced with retroviral vectors containing an intracytoplasmatically truncated version of the human low-affinity nerve growth factor receptor (deltaLNGFR) gene

AU - Fehse, B

AU - Uhde, A

AU - Fehse, Natalja

AU - Eckert, H G

AU - Clausen, Johannes

AU - Rüger, R

AU - Koch, S

AU - Ostertag, W

AU - Zander, A R

AU - Stockschläder, M

PY - 1997/10/10

Y1 - 1997/10/10

N2 - Human hematopoietic stem cells remain one of the most promising target cells for gene therapeutic approaches to treat malignant and nonmalignant diseases. To rapidly characterize transduced cells and to isolate these from residual nontransduced, but biologically equivalent, cells, we have used a Moloney murine leukemia virus (Mo-MuLV)-based retroviral vector containing the intracytoplasmatically truncated human low-affinity nerve growth factor receptor (deltaLNGFR) cDNA as a marker gene. Supernatant transduction of CD34+ cells (mean purity 97%) in fibronectin-coated tissue culture flasks resulted in 5.5-45% (mean 26%) transduced cells expressing deltaLNGFR (LNGFR+ cells). After transduction, more than 65% of the transduced cells remained CD34+. Compared with control (mock- and nontransduced) CD34+ cells, transduction did not decrease the cloning efficiency of CD34+ cells. Immunomagnetic selection of the transduced cells with a monoclonal anti-LNGFR antibody resulted in >90% LNGFR+ cells. Further phenotypic characterization of these highly enriched LNGFR+ cells indicated that the majority co-expressed the CD34 and CD38 antigens. These results show that transduced cells expressing an ectopic cell-surface protein can be rapidly and conveniently quantitated and characterized by fluorescence-activated cell sorting (FACS) analysis and fast and efficiently enriched by immunoadhesion using magnetic beads. The use of cell-surface reporters should facilitate optimization of methods of gene transfer into more primitive hematopoietic progenitors.

AB - Human hematopoietic stem cells remain one of the most promising target cells for gene therapeutic approaches to treat malignant and nonmalignant diseases. To rapidly characterize transduced cells and to isolate these from residual nontransduced, but biologically equivalent, cells, we have used a Moloney murine leukemia virus (Mo-MuLV)-based retroviral vector containing the intracytoplasmatically truncated human low-affinity nerve growth factor receptor (deltaLNGFR) cDNA as a marker gene. Supernatant transduction of CD34+ cells (mean purity 97%) in fibronectin-coated tissue culture flasks resulted in 5.5-45% (mean 26%) transduced cells expressing deltaLNGFR (LNGFR+ cells). After transduction, more than 65% of the transduced cells remained CD34+. Compared with control (mock- and nontransduced) CD34+ cells, transduction did not decrease the cloning efficiency of CD34+ cells. Immunomagnetic selection of the transduced cells with a monoclonal anti-LNGFR antibody resulted in >90% LNGFR+ cells. Further phenotypic characterization of these highly enriched LNGFR+ cells indicated that the majority co-expressed the CD34 and CD38 antigens. These results show that transduced cells expressing an ectopic cell-surface protein can be rapidly and conveniently quantitated and characterized by fluorescence-activated cell sorting (FACS) analysis and fast and efficiently enriched by immunoadhesion using magnetic beads. The use of cell-surface reporters should facilitate optimization of methods of gene transfer into more primitive hematopoietic progenitors.

KW - Animals

KW - Antigens, CD34

KW - Cloning, Molecular

KW - Fibronectins/pharmacology

KW - Gene Transfer Techniques

KW - Genetic Vectors

KW - Hematopoietic Stem Cells/immunology

KW - Humans

KW - Mice

KW - Moloney murine leukemia virus/genetics

KW - Receptors, Nerve Growth Factor/biosynthesis

KW - Time Factors

KW - Transformation, Genetic

U2 - 10.1089/hum.1997.8.15-1815

DO - 10.1089/hum.1997.8.15-1815

M3 - SCORING: Journal article

C2 - 9358031

VL - 8

SP - 1815

EP - 1824

JO - HUM GENE THER

JF - HUM GENE THER

SN - 1043-0342

IS - 15

ER -