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Impact of source tissue and ex vivo expansion on the characterization of goat mesenchymal stem cells
BACKGROUND: There is considerable interest in using goats as models for genetically engineering dairy animals and also for using stem cells as therapeutics for bone and cartilage repair. Mesenchymal stem cells (MSCs) have been isolated and characterized from various species, but are poorly character...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4382838/ https://www.ncbi.nlm.nih.gov/pubmed/25838897 http://dx.doi.org/10.1186/2049-1891-6-1 |
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author | Mohamad-Fauzi, Nuradilla Ross, Pablo J Maga, Elizabeth A Murray, James D |
author_facet | Mohamad-Fauzi, Nuradilla Ross, Pablo J Maga, Elizabeth A Murray, James D |
author_sort | Mohamad-Fauzi, Nuradilla |
collection | PubMed |
description | BACKGROUND: There is considerable interest in using goats as models for genetically engineering dairy animals and also for using stem cells as therapeutics for bone and cartilage repair. Mesenchymal stem cells (MSCs) have been isolated and characterized from various species, but are poorly characterized in goats. RESULTS: Goat MSCs isolated from bone marrow (BM-MSCs) and adipose tissue (ASCs) have the ability to undergo osteogenic, adipogenic and chondrogenic differentiation. Cytochemical staining and gene expression analysis show that ASCs have a greater capacity for adipogenic differentiation compared to BM-MSCs and fibroblasts. Different methods of inducing adipogenesis also affect the extent and profile of adipogenic differentiation in MSCs. Goat fibroblasts were not capable of osteogenesis, hence distinguishing them from the MSCs. Goat MSCs and fibroblasts express CD90, CD105, CD73 but not CD45, and exhibit cytoplasmic localization of OCT4 protein. Goat MSCs can be stably transfected by Nucleofection, but, as evidenced by colony-forming efficiency (CFE), yield significantly different levels of progenitor cells that are robust enough to proliferate into colonies of integrants following G418 selection. BM-MSCs expanded over increasing passages in vitro maintained karyotypic stability up to 20 passages in culture, exhibited an increase in adipogenic differentiation and CFE, but showed altered morphology and amenability to genetic modification by selection. CONCLUSIONS: Our findings provide characterization information on goat MSCs, and show that there can be significant differences between MSCs isolated from different tissues and from within the same tissue. Fibroblasts do not exhibit trilineage differentiation potential at the same capacity as MSCs, making it a more reliable method for distinguishing MSCs from fibroblasts, compared to cell surface marker expression. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/2049-1891-6-1) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-4382838 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-43828382015-04-03 Impact of source tissue and ex vivo expansion on the characterization of goat mesenchymal stem cells Mohamad-Fauzi, Nuradilla Ross, Pablo J Maga, Elizabeth A Murray, James D J Anim Sci Biotechnol Research BACKGROUND: There is considerable interest in using goats as models for genetically engineering dairy animals and also for using stem cells as therapeutics for bone and cartilage repair. Mesenchymal stem cells (MSCs) have been isolated and characterized from various species, but are poorly characterized in goats. RESULTS: Goat MSCs isolated from bone marrow (BM-MSCs) and adipose tissue (ASCs) have the ability to undergo osteogenic, adipogenic and chondrogenic differentiation. Cytochemical staining and gene expression analysis show that ASCs have a greater capacity for adipogenic differentiation compared to BM-MSCs and fibroblasts. Different methods of inducing adipogenesis also affect the extent and profile of adipogenic differentiation in MSCs. Goat fibroblasts were not capable of osteogenesis, hence distinguishing them from the MSCs. Goat MSCs and fibroblasts express CD90, CD105, CD73 but not CD45, and exhibit cytoplasmic localization of OCT4 protein. Goat MSCs can be stably transfected by Nucleofection, but, as evidenced by colony-forming efficiency (CFE), yield significantly different levels of progenitor cells that are robust enough to proliferate into colonies of integrants following G418 selection. BM-MSCs expanded over increasing passages in vitro maintained karyotypic stability up to 20 passages in culture, exhibited an increase in adipogenic differentiation and CFE, but showed altered morphology and amenability to genetic modification by selection. CONCLUSIONS: Our findings provide characterization information on goat MSCs, and show that there can be significant differences between MSCs isolated from different tissues and from within the same tissue. Fibroblasts do not exhibit trilineage differentiation potential at the same capacity as MSCs, making it a more reliable method for distinguishing MSCs from fibroblasts, compared to cell surface marker expression. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/2049-1891-6-1) contains supplementary material, which is available to authorized users. BioMed Central 2015-01-11 /pmc/articles/PMC4382838/ /pubmed/25838897 http://dx.doi.org/10.1186/2049-1891-6-1 Text en © Mohamad-Fauzi et al.; licensee BioMed Central. 2015 This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. 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 Mohamad-Fauzi, Nuradilla Ross, Pablo J Maga, Elizabeth A Murray, James D Impact of source tissue and ex vivo expansion on the characterization of goat mesenchymal stem cells |
title | Impact of source tissue and ex vivo expansion on the characterization of goat mesenchymal stem cells |
title_full | Impact of source tissue and ex vivo expansion on the characterization of goat mesenchymal stem cells |
title_fullStr | Impact of source tissue and ex vivo expansion on the characterization of goat mesenchymal stem cells |
title_full_unstemmed | Impact of source tissue and ex vivo expansion on the characterization of goat mesenchymal stem cells |
title_short | Impact of source tissue and ex vivo expansion on the characterization of goat mesenchymal stem cells |
title_sort | impact of source tissue and ex vivo expansion on the characterization of goat mesenchymal stem cells |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4382838/ https://www.ncbi.nlm.nih.gov/pubmed/25838897 http://dx.doi.org/10.1186/2049-1891-6-1 |
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