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Human Bone Marrow Assessment by Single Cell RNA Sequencing, Mass Cytometry and Flow Cytometry

Karolyn A. Oetjen, Katherine E. Lindblad, Meghali Goswami, Gege Gui, Pradeep K. Dagur, Catherine Lai, Laura W. Dillon, J. Philip McCoy, View ORCID ProfileChristopher S. Hourigan
doi: https://doi.org/10.1101/416750
Karolyn A. Oetjen
Laboratory of Myeloid Malignancies National Heart Lung and Blood Institute, 10 Center Drive, Bethesda, Maryland, USA
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Katherine E. Lindblad
Laboratory of Myeloid Malignancies National Heart Lung and Blood Institute, 10 Center Drive, Bethesda, Maryland, USA
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Meghali Goswami
Laboratory of Myeloid Malignancies National Heart Lung and Blood Institute, 10 Center Drive, Bethesda, Maryland, USA
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Gege Gui
Laboratory of Myeloid Malignancies National Heart Lung and Blood Institute, 10 Center Drive, Bethesda, Maryland, USA
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Pradeep K. Dagur
Flow Cytometry Core, National Heart Lung and Blood Institute, 10 Center Drive, Bethesda, Maryland, USA
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Catherine Lai
Laboratory of Myeloid Malignancies National Heart Lung and Blood Institute, 10 Center Drive, Bethesda, Maryland, USA
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Laura W. Dillon
Laboratory of Myeloid Malignancies National Heart Lung and Blood Institute, 10 Center Drive, Bethesda, Maryland, USA
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J. Philip McCoy
Flow Cytometry Core, National Heart Lung and Blood Institute, 10 Center Drive, Bethesda, Maryland, USA
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Christopher S. Hourigan
Laboratory of Myeloid Malignancies National Heart Lung and Blood Institute, 10 Center Drive, Bethesda, Maryland, USA
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  • ORCID record for Christopher S. Hourigan
  • For correspondence: hourigan@nih.gov
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Abstract

New techniques for single-cell analysis have led to insights into hematopoiesis and the immune system, but the ability of these techniques to cross-validate and reproducibly identify the biological variation in diverse human samples is currently unproven. We therefore performed a comprehensive assessment of human bone marrow cells using both single-cell RNA sequencing and multiparameter flow cytometry from twenty healthy adult human donors across a broad age range. These data characterize variation between healthy donors as well as age-associated changes in cell population frequencies. Direct comparison of techniques revealed discrepancy in the quantification of T lymphocyte and natural killer cell populations. Orthogonal validation of immunophenotyping using mass cytometry demonstrated good correlation with flow cytometry. Technical replicates using single-cell RNA sequencing matched robustly, while biological replicates showed variation. Given the increasing use of single-cell technologies in translational research, this resource serves as an important reference dataset and highlights opportunities for further refinement.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
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Posted September 13, 2018.
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Human Bone Marrow Assessment by Single Cell RNA Sequencing, Mass Cytometry and Flow Cytometry
Karolyn A. Oetjen, Katherine E. Lindblad, Meghali Goswami, Gege Gui, Pradeep K. Dagur, Catherine Lai, Laura W. Dillon, J. Philip McCoy, Christopher S. Hourigan
bioRxiv 416750; doi: https://doi.org/10.1101/416750
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Human Bone Marrow Assessment by Single Cell RNA Sequencing, Mass Cytometry and Flow Cytometry
Karolyn A. Oetjen, Katherine E. Lindblad, Meghali Goswami, Gege Gui, Pradeep K. Dagur, Catherine Lai, Laura W. Dillon, J. Philip McCoy, Christopher S. Hourigan
bioRxiv 416750; doi: https://doi.org/10.1101/416750

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