Arrayed lentiviral barcoding for quantification analysis of hematopoietic dynamics

Authors

  • Jeanne Grosselin,

    1. CEA, Institute of Emerging Diseases and Innovative Therapies (iMETI), F-92265 Fontenay-aux-Roses, Paris, France
    2. Inserm U962 and University Paris Sud 11, CEA-iMETI, F-92265 Fontenay-aux-Roses, Paris, France
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    • Authors contributions: J.G.: conception and design, collection and assembly of data, and data analysis and interpretation; K.S.F.: conception and design, collection and assembly of data, data analysis and interpretation, manuscript writing, and final approval of manuscript; E.P.: conception and design and data analysis and interpretation; S.C.: data analysis and interpretation and administrative support; D.T.L.R.: conception and design, data analysis and interpretation, and manuscript writing; P.L.: conception and design, data analysis and interpretation, manuscript writing, and final approval of manuscript. J.G., K.S.F., D.T.L.R., and P.L. contributed equally to this article.

  • Karine Sii-Felice,

    Corresponding author
    1. CEA, Institute of Emerging Diseases and Innovative Therapies (iMETI), F-92265 Fontenay-aux-Roses, Paris, France
    2. Inserm U962 and University Paris Sud 11, CEA-iMETI, F-92265 Fontenay-aux-Roses, Paris, France
    • M.D., CEA, Institute of Emerging Diseases and Innovative Therapies (iMETI), 18, route du Panorama BP-6, 92265 Fontenay-aux-Roses, Paris, FrancePh.D., CEA, Institute of Emerging Diseases and Innovative Therapies (iMETI), 18, route du Panorama BP-6, 92265 Fontenay-aux-Roses, Paris, France. E-mail: karine.sii-felice@cea.fr pleboulch@rics.bwh.harvard.edu Telephone: +33-(0)1-46-54-70-55; Fax: +33-(0)1-46-54-74-99 Telephone: +33-(0)1-46-54-83-49; Fax: +33-(0)1-46-54-74-99

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  • Emmanuel Payen,

    1. CEA, Institute of Emerging Diseases and Innovative Therapies (iMETI), F-92265 Fontenay-aux-Roses, Paris, France
    2. Inserm U962 and University Paris Sud 11, CEA-iMETI, F-92265 Fontenay-aux-Roses, Paris, France
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    • Authors contributions: J.G.: conception and design, collection and assembly of data, and data analysis and interpretation; K.S.F.: conception and design, collection and assembly of data, data analysis and interpretation, manuscript writing, and final approval of manuscript; E.P.: conception and design and data analysis and interpretation; S.C.: data analysis and interpretation and administrative support; D.T.L.R.: conception and design, data analysis and interpretation, and manuscript writing; P.L.: conception and design, data analysis and interpretation, manuscript writing, and final approval of manuscript. J.G., K.S.F., D.T.L.R., and P.L. contributed equally to this article.

  • Stany Chretien,

    1. CEA, Institute of Emerging Diseases and Innovative Therapies (iMETI), F-92265 Fontenay-aux-Roses, Paris, France
    2. Inserm U962 and University Paris Sud 11, CEA-iMETI, F-92265 Fontenay-aux-Roses, Paris, France
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    • Authors contributions: J.G.: conception and design, collection and assembly of data, and data analysis and interpretation; K.S.F.: conception and design, collection and assembly of data, data analysis and interpretation, manuscript writing, and final approval of manuscript; E.P.: conception and design and data analysis and interpretation; S.C.: data analysis and interpretation and administrative support; D.T.L.R.: conception and design, data analysis and interpretation, and manuscript writing; P.L.: conception and design, data analysis and interpretation, manuscript writing, and final approval of manuscript. J.G., K.S.F., D.T.L.R., and P.L. contributed equally to this article.

  • Diana Tronik-Le Roux,

    1. CEA, Institute of Emerging Diseases and Innovative Therapies (iMETI), F-92265 Fontenay-aux-Roses, Paris, France
    2. Inserm U962 and University Paris Sud 11, CEA-iMETI, F-92265 Fontenay-aux-Roses, Paris, France
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    • Authors contributions: J.G.: conception and design, collection and assembly of data, and data analysis and interpretation; K.S.F.: conception and design, collection and assembly of data, data analysis and interpretation, manuscript writing, and final approval of manuscript; E.P.: conception and design and data analysis and interpretation; S.C.: data analysis and interpretation and administrative support; D.T.L.R.: conception and design, data analysis and interpretation, and manuscript writing; P.L.: conception and design, data analysis and interpretation, manuscript writing, and final approval of manuscript. J.G., K.S.F., D.T.L.R., and P.L. contributed equally to this article.

  • Philippe Leboulch

    Corresponding author
    1. CEA, Institute of Emerging Diseases and Innovative Therapies (iMETI), F-92265 Fontenay-aux-Roses, Paris, France
    2. Inserm U962 and University Paris Sud 11, CEA-iMETI, F-92265 Fontenay-aux-Roses, Paris, France
    3. Genetics Division, Brigham & Women's Hospital and Harvard Medical School, Boston, Massachusetts, USA
    • M.D., CEA, Institute of Emerging Diseases and Innovative Therapies (iMETI), 18, route du Panorama BP-6, 92265 Fontenay-aux-Roses, Paris, FrancePh.D., CEA, Institute of Emerging Diseases and Innovative Therapies (iMETI), 18, route du Panorama BP-6, 92265 Fontenay-aux-Roses, Paris, France. E-mail: karine.sii-felice@cea.fr pleboulch@rics.bwh.harvard.edu Telephone: +33-(0)1-46-54-70-55; Fax: +33-(0)1-46-54-74-99 Telephone: +33-(0)1-46-54-83-49; Fax: +33-(0)1-46-54-74-99

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Abstract

Our understanding of system dynamics of mixed cell populations in whole organisms has benefited from the advent of individual cell marking by nonarrayed DNA barcodes subsequently analyzed by high-throughput DNA sequencing. However, key limitations include statistical biases compromising quantification and the lack of applicability to deconvolute individual cell fate in vivo after pooling single cells differentially exposed to different conditions ex vivo. Here, we have derived an arrayed lentiviral library of DNA barcodes and obtained a proof-of-concept of its resolving capacity by quantifying hematopoietic regeneration after engraftment of mice with genetically modified autologous cells. This method has helped clarify and bridge the seemingly opposed clonal-succession and continuous-recruitment models of hematopoietic stem cell behavior and revealed that myeloid-lymphoid biases are common occurrences in steady-state hematopoiesis. Arrayed lentiviral barcoding should prove a versatile and powerful approach to deconvolute cell dynamics in vivo with applications in hematology, embryology, and cancer biology. STEM Cells 2013;31:2162–2171

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