Measurement of generation-dependent proliferation rates and death rates during mouse erythroid progenitor cell differentiation

Authors

  • Vahe Akbarian,

    1. Institute of Biomaterials and Biomedical Engineering (IBBME), University of Toronto, Toronto, Canada
    2. Donnelly Centre, University of Toronto, Toronto, Canada
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    • V. Akbarian and W. Wang contributed equally to the manuscript.

  • Weijia Wang,

    1. Institute of Biomaterials and Biomedical Engineering (IBBME), University of Toronto, Toronto, Canada
    2. Donnelly Centre, University of Toronto, Toronto, Canada
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    • V. Akbarian and W. Wang contributed equally to the manuscript.

  • Julie Audet

    Corresponding author
    1. Institute of Biomaterials and Biomedical Engineering (IBBME), University of Toronto, Toronto, Canada
    2. Donnelly Centre, University of Toronto, Toronto, Canada
    • Ph.D., Associate Professor, Institute of Biomaterials and Biomedical Engineering, University of Toronto, 164 College Street, Rm 407, Toronto, ON, Canada, M5S 3G9
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Abstract

Herein, we describe an experimental and computational approach to perform quantitative carboxyfluorescein diacetate succinimidyl ester (CFSE) cell-division tracking in cultures of primary colony-forming unit-erythroid (CFU-E) cells, a hematopoietic progenitor cell type, which is an important target for the treatment of blood disorders and for the manufacture of red blood cells. CFSE labeling of CFU-Es isolated from mouse fetal livers was performed to examine the effects of stem cell factor (SCF) and erythropoietin (EPO) in culture. We used a dynamic model of proliferation based on the Smith-Martin representation of the cell cycle to extract proliferation rates and death rates from CFSE time-series. However, we found that to accurately represent the cell population dynamics in differentiation cultures of CFU-Es, it was necessary to develop a model with generation-specific rate parameters. The generation-specific rates of proliferation and death were extracted for six generations (G0G5) and they revealed that, although SCF alone or EPO alone supported similar total cell outputs in culture, stimulation with EPO resulted in significantly higher proliferation rates from G2 to G5 and higher death rates in G2, G3, and G5 compared with SCF. In addition, proliferation rates tended to increase from G1 to G5 in cultures supplemented with EPO and EPO + SCF, while they remained lower and more constant across generations with SCF. The results are consistent with the notion that SCF promotes CFU-E self-renewal while EPO promotes CFU-E differentiation in culture. © 2012 International Society for Advancement of Cytometry

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