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

Abstract

Both stem cell chemokine stromal cell-derived factor-1α (SDF-1α) and extracellular nucleotides such as adenosine triphosphate (ATP) are increased in ischemic myocardium. Since ATP has been reported to influence cell migration, we analysed the migratory response of bone marrow cells towards a combination of SDF-1 and ATP. Total nucleated cells (BM-TNCs) were isolated from bone marrow of cardiac surgery patients. Migration assays were performed in vitro. Subsequently, migrated cells were subjected to multicolor flow cytometric analysis of CD133, CD34, CD117, CD184, CD309, and CD14 expression. BM-TNCs migrated significantly towards a combination of SDF-1 and ATP. The proportions of CD34+ cells as well as subpopulations coexpressing multiple stem cell markers were selectively enhanced after migration towards SDF-1 or SDF-1 + ATP. After spontaneous migration, significantly fewer stem cells and CD184+ cells were detected. Direct incubation with SDF-1 led to a reduction of CD184+ but not stem cell marker-positive cells, while incubation with ATP significantly increased CD14+ percentage. In summary, we found that while a combination of SDF-1 and ATP elicited strong migration of BM-TNCs in vitro, only SDF-1 was responsible for selective attraction of hematopoietic stem cells. Meanwhile, spontaneous migration of stem cells was lower compared to BM-TNCs or monocytes.

H2O2‐induced oxidative stress disrupts mitochondrial functions and impairs migratory potential of human epidermal melanocytes

“…*P < 0.05, vs control group upstream regulator for mitochondrial ATP production. [36] A similar effect was also recently observed in keratinocytes in vitro. [37] The maintenance of normal morphology, mitochondrial permeability and ATP release are prerequisite factors for normal mitochondrial function.…”

Mechanisms of stem cell based cardiac repair-gap junctional signaling promotes the cardiac lineage specification of mesenchymal stem cells

“…Next, we investigated the quality of MACS-based cell isolation by evaluating purity and viability of each cell fraction. Isolated CD133 + cells were analyzed using an adapted ISHAGE gating strategy for CD34 + /CD133 + cells 22 . For analysis of the CD271 + fraction, cells were stained for CD271 and CD45 surface markers.…”

GMP-conformant on-site manufacturing of a CD133+ stem cell product for cardiovascular regeneration

“…We assume that within this non-target population one fraction (CD45 + /CD34 low/+ /CD133 − ) represents hematopoietic stem and progenitor cells [ 35 ]. The second non-target fraction (CD45 + /CD34 − /CD133 − ) seems to be in a more progressed stage of maturation as indicated by a lower expression of the stem cell marker CD117 and a higher expression of the maturation marker CD14 [ 26 ]. Moreover, this fraction shows the highest expression of CD184 (CXCR4) as well as CD309 (KDR, VEGFR2) indicating a possible role in stem cell homing and in the regulation of angiogenesis, respectively [ 30 , 36 , 37 ].…”

“…Compensation was established using single-stained controls and gating was performed with matched isotype/fluorescence minus one (FMO) controls. The Boolean gating strategy for CD133 + cells was arranged on the basis of the International Society of Hematotherapy and Graft Engineering (ISHAGE) guidelines for CD34 + cell analysis [ 25 ] as described previously by our group [ 26 ].…”

“…The expression of additional stemness (CD117, CD184, CD309) and maturation (CD14) markers in the CP was analyzed by flow cytometry in a multicolor panel as described previously by our group [ 26 ]. Data acquisition was performed based on ISHAGE gating strategy used for QC.…”