Efficient adenoviral vector transduction of human hematopoietic SCID-repopulating and long-term culture-initiating cells

This article presents our studies on the adenoviral transduction efficiency, level of transgene expression, cell cycle status, and multilineage reconstitution ability of human CD34+ hematopoietic cells transduced under proliferating and survival growth conditions. Bone marrow and umbilical cord bloo...

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Veröffentlicht in:Human gene therapy 2000-06, Vol.11 (9), p.1313-1327
Hauptverfasser: XIAOLONG FAN, BRUN, A, SEGREN, S, JACOBSEN, S. E. W, KARLSSON, S
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container_end_page 1327
container_issue 9
container_start_page 1313
container_title Human gene therapy
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creator XIAOLONG FAN
BRUN, A
SEGREN, S
JACOBSEN, S. E. W
KARLSSON, S
description This article presents our studies on the adenoviral transduction efficiency, level of transgene expression, cell cycle status, and multilineage reconstitution ability of human CD34+ hematopoietic cells transduced under proliferating and survival growth conditions. Bone marrow and umbilical cord blood CD34+ cells were cultured in serum-free medium under survival conditions with thrombopoietin (Tpo) alone, or under proliferating conditions with Tpo, c-Kit ligand (KL), and Flt3 ligand (FL). Adenoviral vectors carrying the enhanced green fluorescent protein (EGFP) gene under the control of the PGK-1 promoter were used to transduce CD34+ cells. Approximately 10% of CD34+ cells were EGFP+ under both culture conditions. In contrast, up to 50% of CD34+CD38- cells were EGFP+, whereas a maximum of 8% of CD34+CD38(high) cells were EGFP+ (p < 0.001). Both colony-forming unit cells (CFU-C) and 5-week long-term culture-initiating cells (LTC-ICs) were efficiently transduced. Under survival conditions, a substantial fraction of transduced CD34+ cells remained quiescent. The nondividing CD34+EGFP+ cells contained LTC-ICs capable of reconstituting longterm culture for as long as 10 weeks. CD34+EGFP+ cells also retained the ability to engraft and multilineage-reconstitute NOD/SCID mice. These observations demonstrate that primitive human hematopoietic progenitor cells can be efficiently transduced by adenoviral vectors.
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Both colony-forming unit cells (CFU-C) and 5-week long-term culture-initiating cells (LTC-ICs) were efficiently transduced. Under survival conditions, a substantial fraction of transduced CD34+ cells remained quiescent. The nondividing CD34+EGFP+ cells contained LTC-ICs capable of reconstituting longterm culture for as long as 10 weeks. CD34+EGFP+ cells also retained the ability to engraft and multilineage-reconstitute NOD/SCID mice. 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E. W</creatorcontrib><creatorcontrib>KARLSSON, S</creatorcontrib><title>Efficient adenoviral vector transduction of human hematopoietic SCID-repopulating and long-term culture-initiating cells</title><title>Human gene therapy</title><addtitle>Hum Gene Ther</addtitle><description>This article presents our studies on the adenoviral transduction efficiency, level of transgene expression, cell cycle status, and multilineage reconstitution ability of human CD34+ hematopoietic cells transduced under proliferating and survival growth conditions. Bone marrow and umbilical cord blood CD34+ cells were cultured in serum-free medium under survival conditions with thrombopoietin (Tpo) alone, or under proliferating conditions with Tpo, c-Kit ligand (KL), and Flt3 ligand (FL). Adenoviral vectors carrying the enhanced green fluorescent protein (EGFP) gene under the control of the PGK-1 promoter were used to transduce CD34+ cells. 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W ; KARLSSON, S</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c357t-659a43e53b1486709babc9903efb3106d2cc3c99485ab157fc6d77c67c5d141a3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2000</creationdate><topic>Adenoviridae</topic><topic>Adenovirus</topic><topic>ADP-ribosyl Cyclase</topic><topic>ADP-ribosyl Cyclase 1</topic><topic>Animals</topic><topic>Antigens, CD</topic><topic>Antigens, CD34 - analysis</topic><topic>Antigens, Differentiation - analysis</topic><topic>Biological and medical sciences</topic><topic>Biotechnology</topic><topic>Bone Marrow Cells - cytology</topic><topic>CD34 antigen</topic><topic>Cell Division</topic><topic>Cell Lineage</topic><topic>Cell Survival</topic><topic>Fetal Blood - cytology</topic><topic>Fundamental and applied biological sciences. 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Economical aspects</topic><topic>Luminescent Proteins - genetics</topic><topic>Membrane Glycoproteins</topic><topic>Membrane Proteins - physiology</topic><topic>Mice</topic><topic>Mice, SCID</topic><topic>NAD+ Nucleosidase - analysis</topic><topic>Stem Cell Factor - physiology</topic><topic>Thrombopoietin - physiology</topic><topic>Transduction, Genetic</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>XIAOLONG FAN</creatorcontrib><creatorcontrib>BRUN, A</creatorcontrib><creatorcontrib>SEGREN, S</creatorcontrib><creatorcontrib>JACOBSEN, S. E. 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Bone marrow and umbilical cord blood CD34+ cells were cultured in serum-free medium under survival conditions with thrombopoietin (Tpo) alone, or under proliferating conditions with Tpo, c-Kit ligand (KL), and Flt3 ligand (FL). Adenoviral vectors carrying the enhanced green fluorescent protein (EGFP) gene under the control of the PGK-1 promoter were used to transduce CD34+ cells. Approximately 10% of CD34+ cells were EGFP+ under both culture conditions. In contrast, up to 50% of CD34+CD38- cells were EGFP+, whereas a maximum of 8% of CD34+CD38(high) cells were EGFP+ (p &lt; 0.001). Both colony-forming unit cells (CFU-C) and 5-week long-term culture-initiating cells (LTC-ICs) were efficiently transduced. Under survival conditions, a substantial fraction of transduced CD34+ cells remained quiescent. The nondividing CD34+EGFP+ cells contained LTC-ICs capable of reconstituting longterm culture for as long as 10 weeks. CD34+EGFP+ cells also retained the ability to engraft and multilineage-reconstitute NOD/SCID mice. These observations demonstrate that primitive human hematopoietic progenitor cells can be efficiently transduced by adenoviral vectors.</abstract><cop>Larchmont, NY</cop><pub>Liebert</pub><pmid>10890741</pmid><doi>10.1089/10430340050032410</doi><tpages>15</tpages></addata></record>
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identifier ISSN: 1043-0342
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subjects Adenoviridae
Adenovirus
ADP-ribosyl Cyclase
ADP-ribosyl Cyclase 1
Animals
Antigens, CD
Antigens, CD34 - analysis
Antigens, Differentiation - analysis
Biological and medical sciences
Biotechnology
Bone Marrow Cells - cytology
CD34 antigen
Cell Division
Cell Lineage
Cell Survival
Fetal Blood - cytology
Fundamental and applied biological sciences. Psychology
Gene Expression
Gene therapy
Genetic Vectors - genetics
green fluorescent protein
Green Fluorescent Proteins
Health. Pharmaceutical industry
Hematopoietic Stem Cell Transplantation
Hematopoietic Stem Cells - metabolism
Humans
Industrial applications and implications. Economical aspects
Luminescent Proteins - genetics
Membrane Glycoproteins
Membrane Proteins - physiology
Mice
Mice, SCID
NAD+ Nucleosidase - analysis
Stem Cell Factor - physiology
Thrombopoietin - physiology
Transduction, Genetic
title Efficient adenoviral vector transduction of human hematopoietic SCID-repopulating and long-term culture-initiating cells
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