Abstract
BackgroundThe number of Mesenchymal Stem/Stromal Cells (MSCs) in the human bone marrow (BM) is small compared to other cell types. BM aspirate concentration (BMAC) may be used to increase numbers of MSCs, but the composition of MSC subpopulations and growth factors after processing are unknown. The purpose of this study was to assess the enrichment of stem/progenitor cells and growth factors in BM aspirate by two different commercial concentration devices versus standard BM aspiration.Methods120 mL of BM was aspirated from the iliac crest of 10 male donors. Each sample was processed simultaneously by either Emcyte GenesisCS® (Emcyte) or Harvest SmartPReP2 BMAC (Harvest) devices and compared to untreated BM aspirate. Samples were analyzed with multicolor flow cytometry for cellular viability and expression of stem/progenitor cells markers. Stem/progenitor cell content was verified by quantification of colony forming unit-fibroblasts (CFU-F). Platelet, red blood cell and total nucleated cell (TNC) content were determined using an automated hematology analyzer. Growth factors contents were analyzed with protein quantification assays. Statistical analyses were performed by ANOVA analysis of variance followed by Tukey’s multiple comparison test or Wilcoxon matched-pairs signed rank test with p < 0.05 for significance.ResultsCell viability after processing was approximately 90% in all groups. Compared to control, both devices significantly enriched TNCs and platelets, as well as the CD45−CD73+ and CD45−CD73+CD90+ cell populations. Further, Harvest significantly concentrated CD45−CD10+, CD45−CD29+, CD45−CD90+, CD45−CD105+, CD45−CD119+ cells, and CD45dimCD90+CD271+ MSCs, whereas Emcyte significantly enriched CD45dimCD44+CD271+ MSCs. BM concentration also increased the numbers of CFU-F, platelet-derived growth factor, vascular endothelial growth factor, macrophage colony-stimulating factor, interleukin-1b, VCAM-1 and total protein. Neither system concentrated red blood cells, hematopoietic stem cells or bone morphogenetic proteins.ConclusionThis data could contribute to the development of BMAC quality control assays as both BMAC systems concentrated platelets, growth factors and non-hematopoietic stem cell subpopulations with distinct phenotypes without loss of cell viability when compared to unprocessed BM.
Highlights
The number of Mesenchymal Stem/Stromal Cells (MSCs) in the human bone marrow (BM) is small compared to other cell types
Concentrations of total nucleated cell (TNC) were significantly higher in both BM aspirate concentration (BMAC) groups compared to controls indicating an approx. tenfold concentration (Harvest: 235.11 × 103/ μL, p = 0.0472; Emcyte: 265.28 × 103/μL, p = 0.0307; control: 24.49 × 103/μL) (Fig. 2a)
The content of distinct CD45+ cell populations was enriched after BM aspirate concentration, i.e. CD45+73+ (Harvest: 2.71 fold vs control, p = 0.0313; Emcyte: 2.13 vs control, p = 0.1563), CD45+90+ (Harvest: 2.61 fold vs control, p = 0.0781; Emcyte: 2.74 vs control, p = 0.0469), Fig. 2 Blood cell counts in both BMAC groups and controls
Summary
The number of Mesenchymal Stem/Stromal Cells (MSCs) in the human bone marrow (BM) is small compared to other cell types. BM aspirate concentration (BMAC) may be used to increase numbers of MSCs, but the composition of MSC subpopulations and growth factors after processing are unknown. The purpose of this study was to assess the enrichment of stem/progenitor cells and growth factors in BM aspirate by two different commercial concentration devices versus standard BM aspiration. BMAC is obtained through density gradient centrifugation of bone marrow (BM) typically aspirated from the iliac crest It is currently approved by the United States Food and Drug Administration for clinical use as a means of acquiring progenitor cells and growth factors to promote healing in the orthopaedic patient [2]. It is imperative to compare BMAC produced by commercially available systems to better understand its therapeutic potential and to eventually establish a correlation between the progenitor cell composition, the concentration of biologic factors and clinical outcome
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