Quantitation of progenitor cell populations and growth factors after bone marrow aspirate concentration
Abstract Background 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 unk...
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doaj-60d47976869149f2b8f9e18ef747ff9f2020-11-25T02:03:06ZengBMCJournal of Translational Medicine1479-58762019-04-011711910.1186/s12967-019-1866-7Quantitation of progenitor cell populations and growth factors after bone marrow aspirate concentrationRichard Schäfer0Malcolm R. DeBaun1Erika Fleck2Christopher J. Centeno3Daniela Kraft4Johannes Leibacher5Karen Bieback6Erhard Seifried7Jason L. Dragoo8Institute for Transfusion Medicine and Immunohematology, German Red Cross Blood Service Baden-Württemberg – Hessen gGmbHDepartment of Orthopedic Surgery, Stanford University School of MedicineInstitute for Transfusion Medicine and Immunohematology, German Red Cross Blood Service Baden-Württemberg – Hessen gGmbHCenteno-Schultz ClinicInstitute for Transfusion Medicine and Immunohematology, German Red Cross Blood Service Baden-Württemberg – Hessen gGmbHInstitute for Transfusion Medicine and Immunohematology, German Red Cross Blood Service Baden-Württemberg – Hessen gGmbHInstitute of Transfusion Medicine and Immunology, Medical Faculty Mannheim, Heidelberg University, German Red Cross Blood Service Baden-Württemberg – Hessen gGmbHInstitute for Transfusion Medicine and Immunohematology, German Red Cross Blood Service Baden-Württemberg – Hessen gGmbHDepartment of Orthopedic Surgery, Stanford University School of MedicineAbstract Background 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. Methods 120 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. Results Cell 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. Conclusion This 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.http://link.springer.com/article/10.1186/s12967-019-1866-7Bone marrow aspirate concentrateBone marrow aspirationBMACGrowth factorsMesenchymal stem cellStem cell |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Richard Schäfer Malcolm R. DeBaun Erika Fleck Christopher J. Centeno Daniela Kraft Johannes Leibacher Karen Bieback Erhard Seifried Jason L. Dragoo |
spellingShingle |
Richard Schäfer Malcolm R. DeBaun Erika Fleck Christopher J. Centeno Daniela Kraft Johannes Leibacher Karen Bieback Erhard Seifried Jason L. Dragoo Quantitation of progenitor cell populations and growth factors after bone marrow aspirate concentration Journal of Translational Medicine Bone marrow aspirate concentrate Bone marrow aspiration BMAC Growth factors Mesenchymal stem cell Stem cell |
author_facet |
Richard Schäfer Malcolm R. DeBaun Erika Fleck Christopher J. Centeno Daniela Kraft Johannes Leibacher Karen Bieback Erhard Seifried Jason L. Dragoo |
author_sort |
Richard Schäfer |
title |
Quantitation of progenitor cell populations and growth factors after bone marrow aspirate concentration |
title_short |
Quantitation of progenitor cell populations and growth factors after bone marrow aspirate concentration |
title_full |
Quantitation of progenitor cell populations and growth factors after bone marrow aspirate concentration |
title_fullStr |
Quantitation of progenitor cell populations and growth factors after bone marrow aspirate concentration |
title_full_unstemmed |
Quantitation of progenitor cell populations and growth factors after bone marrow aspirate concentration |
title_sort |
quantitation of progenitor cell populations and growth factors after bone marrow aspirate concentration |
publisher |
BMC |
series |
Journal of Translational Medicine |
issn |
1479-5876 |
publishDate |
2019-04-01 |
description |
Abstract Background 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. Methods 120 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. Results Cell 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. Conclusion This 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. |
topic |
Bone marrow aspirate concentrate Bone marrow aspiration BMAC Growth factors Mesenchymal stem cell Stem cell |
url |
http://link.springer.com/article/10.1186/s12967-019-1866-7 |
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