Selective Migration of Subpopulations of Bone Marrow Cells along an SDF-1α and ATP Gradient

Figure 1

Flow cytometric gating strategy. Debris was excluded in a forward/side scatter (FSC/SSC) dot plot (a); viable cells were defined as cells with low/negative fluorescence in the APC-Cy7 channel (b). Hematopoietic stem cells were selected with an adapted ISHAGE gating strategy [25], exemplified for CD34 in (c)–(e): cells positive for CD34-FITC were gated (c); lymphocytes were excluded based on their high expression of CD45 (d). A FSC/SSC backgate was employed to select CD34+ cells with blast morphology (e). CD133+ and CD117+ HSCs were analysed accordingly ((f) and (g), resp.). CD184+ (CXCR4+), CD309+ (KDR+), and CD14+ cells were gated on viable cells ((h), (i), and (j), resp.). CD34+ CD133+ double positive cell population was gated in a FITC/PE plot (k). Subpopulations coexpressing further markers were selected from the ISHAGE-gated stem cell populations. CD34+ CD133+ CD117+ cells were analysed from ISHAGE-gated CD34+ CD133+ cells, and CD34+ CD117+ cells were analysed based on ISHAGE CD34+ cells.

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