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Quantitation of progenitor cell populations and growth factors after bone marrow aspirate concentration
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. Th...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6454687/ https://www.ncbi.nlm.nih.gov/pubmed/30961655 http://dx.doi.org/10.1186/s12967-019-1866-7 |
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author | Schäfer, Richard DeBaun, Malcolm R. Fleck, Erika Centeno, Christopher J. Kraft, Daniela Leibacher, Johannes Bieback, Karen Seifried, Erhard Dragoo, Jason L. |
author_facet | Schäfer, Richard DeBaun, Malcolm R. Fleck, Erika Centeno, Christopher J. Kraft, Daniela Leibacher, Johannes Bieback, Karen Seifried, Erhard Dragoo, Jason L. |
author_sort | Schäfer, Richard |
collection | PubMed |
description | 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. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12967-019-1866-7) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-6454687 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-64546872019-04-19 Quantitation of progenitor cell populations and growth factors after bone marrow aspirate concentration Schäfer, Richard DeBaun, Malcolm R. Fleck, Erika Centeno, Christopher J. Kraft, Daniela Leibacher, Johannes Bieback, Karen Seifried, Erhard Dragoo, Jason L. J Transl Med Research 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. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12967-019-1866-7) contains supplementary material, which is available to authorized users. BioMed Central 2019-04-08 /pmc/articles/PMC6454687/ /pubmed/30961655 http://dx.doi.org/10.1186/s12967-019-1866-7 Text en © The Author(s) 2019 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. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Schäfer, Richard DeBaun, Malcolm R. Fleck, Erika Centeno, Christopher J. Kraft, Daniela Leibacher, Johannes Bieback, Karen Seifried, Erhard Dragoo, Jason L. Quantitation of progenitor cell populations and growth factors after bone marrow aspirate concentration |
title | 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_short | 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 |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6454687/ https://www.ncbi.nlm.nih.gov/pubmed/30961655 http://dx.doi.org/10.1186/s12967-019-1866-7 |
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