Clonal analysis of individual marrow-repopulating cells after experimental peripheral blood progenitor cell transplantion

K. Zsuzsanna Nagy; Stephanie Laufs; Bernhard Gentner; Sonja Naundorf; Klaus Kuehlcke; Julian Topaly; Eike C. Buss; W. Jens Zeller; Stefan Fruehauf

Clonal analysis of individual marrow-repopulating cells after experimental peripheral blood progenitor cell transplantion

K. Zsuzsanna Nagy, Stephanie Laufs, Bernhard Gentner, Sonja Naundorf, Klaus Kuehlcke, Julian Topaly, Eike C. Buss, W. Jens Zeller, Stefan Fruehauf

IRCCS Ospedale San Raffaele

Research output: Contribution to journal › Article › peer-review

Abstract

Methods to analyze the clonality of an adverse event in preclinical or clinical retroviral stem cell gene therapy protocols are needed. We analyzed the progeny of retrovirally transduced human peripheral blood progenitor cells (PBPCs) after transplantation and engraftment in immune-deficient mice. The integration site of the provirus serves as a unique tag of the individual transduced PBPC. A plasmid library of junctions between proviral and human genomic DNA was generated. We were able to detect individual transduced cell clones that amounted to 0.14%-0.0001% of chimeric bone marrow cells. This is the first report in which the contribution of individual marrow-repopulating cells to human hematopoiesis is directly quantified.

Original language	English
Pages (from-to)	570-579
Number of pages	10
Journal	Stem Cells
Volume	22
Issue number	4
Publication status	Published - 2004

Keywords

Experimental PBPC transplantation
SCID-repopulating cells
Single cell analysis

ASJC Scopus subject areas

Cell Biology

Cite this

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abstract = "Methods to analyze the clonality of an adverse event in preclinical or clinical retroviral stem cell gene therapy protocols are needed. We analyzed the progeny of retrovirally transduced human peripheral blood progenitor cells (PBPCs) after transplantation and engraftment in immune-deficient mice. The integration site of the provirus serves as a unique tag of the individual transduced PBPC. A plasmid library of junctions between proviral and human genomic DNA was generated. We were able to detect individual transduced cell clones that amounted to 0.14%-0.0001% of chimeric bone marrow cells. This is the first report in which the contribution of individual marrow-repopulating cells to human hematopoiesis is directly quantified.",

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AU - Nagy, K. Zsuzsanna

AU - Laufs, Stephanie

AU - Gentner, Bernhard

AU - Naundorf, Sonja

AU - Kuehlcke, Klaus

AU - Topaly, Julian

AU - Buss, Eike C.

AU - Zeller, W. Jens

AU - Fruehauf, Stefan

PY - 2004

Y1 - 2004

N2 - Methods to analyze the clonality of an adverse event in preclinical or clinical retroviral stem cell gene therapy protocols are needed. We analyzed the progeny of retrovirally transduced human peripheral blood progenitor cells (PBPCs) after transplantation and engraftment in immune-deficient mice. The integration site of the provirus serves as a unique tag of the individual transduced PBPC. A plasmid library of junctions between proviral and human genomic DNA was generated. We were able to detect individual transduced cell clones that amounted to 0.14%-0.0001% of chimeric bone marrow cells. This is the first report in which the contribution of individual marrow-repopulating cells to human hematopoiesis is directly quantified.

AB - Methods to analyze the clonality of an adverse event in preclinical or clinical retroviral stem cell gene therapy protocols are needed. We analyzed the progeny of retrovirally transduced human peripheral blood progenitor cells (PBPCs) after transplantation and engraftment in immune-deficient mice. The integration site of the provirus serves as a unique tag of the individual transduced PBPC. A plasmid library of junctions between proviral and human genomic DNA was generated. We were able to detect individual transduced cell clones that amounted to 0.14%-0.0001% of chimeric bone marrow cells. This is the first report in which the contribution of individual marrow-repopulating cells to human hematopoiesis is directly quantified.

KW - Experimental PBPC transplantation

KW - SCID-repopulating cells

KW - Single cell analysis

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Clonal analysis of individual marrow-repopulating cells after experimental peripheral blood progenitor cell transplantion

Abstract

Keywords

ASJC Scopus subject areas

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