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. 2016 Apr 7;5:15. doi: 10.1186/s40169-016-0092-7

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© Pule et al. 2016

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

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Fig. 1 — Flow cytometry showing differentiation of CD34⁺ cells to erythroid progenitors (a) and the cluster-like basophilic erythroblast morphology of erythroid cells on day 7 of differentiation (b) and the colony-forming hematopoietic cell units (CFU-E) (c). CD34⁺ hematopoietic stem cells were purified from umbilical cord blood mononuclear cells using Dynabead magnetic separation to a purity of 87.6 %. After the 15 day single-phase ex vivo expansion and differentiation, the cell population was 92.7 and 78.2 % CD235a⁺ and CD71⁺, respectively with minimal CD34 positivity. This confirmed our differentiation and provided erythroid cells for down-stream experiments (a). During the expansion and differentiation of HSCs to erythroid cells, the cells formed cluster-like colonies before separating into single cells in suspension, which was typical of terminal differentiation (b). The hematopoietic colony formation assay was performed to confirm the stem-like behaviour of the Dyna-bead selected CD34⁺ HSCs prior to expansion and differentiation. The colony-forming unit- granulocyte, erythroid, monocyte, megakaryocyte (CFU-GEMM) semi-solid cultures were grown for 14 days and stained for colony counting (c)