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Functional properties of equine adipose-derived mesenchymal stromal cells cultured with equine platelet lysate
Successful translation of multipotent mesenchymal stromal cell (MSC)-based therapies into clinical reality relies on adequate cell production procedures. These should be available not only for human MSC, but also for MSC from animal species relevant to preclinical research and veterinary medicine. T...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9395693/ https://www.ncbi.nlm.nih.gov/pubmed/36016806 http://dx.doi.org/10.3389/fvets.2022.890302 |
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author | Hagen, Alina Niebert, Sabine Brandt, Vivian-Pascal Holland, Heidrun Melzer, Michaela Wehrend, Axel Burk, Janina |
author_facet | Hagen, Alina Niebert, Sabine Brandt, Vivian-Pascal Holland, Heidrun Melzer, Michaela Wehrend, Axel Burk, Janina |
author_sort | Hagen, Alina |
collection | PubMed |
description | Successful translation of multipotent mesenchymal stromal cell (MSC)-based therapies into clinical reality relies on adequate cell production procedures. These should be available not only for human MSC, but also for MSC from animal species relevant to preclinical research and veterinary medicine. The cell culture medium supplementation is one of the critical aspects in MSC production. Therefore, we previously established a scalable protocol for the production of buffy-coat based equine platelet lysate (ePL). This ePL proved to be a suitable alternative to fetal bovine serum (FBS) for equine adipose-derived (AD-) MSC culture so far, as it supported AD-MSC proliferation and basic characteristics. The aim of the current study was to further analyze the functional properties of equine AD-MSC cultured with the same ePL, focusing on cell fitness, genetic stability and pro-angiogenic potency. All experiments were performed with AD-MSC from n = 5 horses, which were cultured either in medium supplemented with 10% FBS, 10% ePL or 2.5% ePL. AD-MSC cultured with 2.5% ePL, which previously showed decreased proliferation potential, displayed higher apoptosis but lower senescence levels as compared to 10% ePL medium (p < 0.05). Non-clonal chromosomal aberrations occurred in 8% of equine AD-MSC cultivated with FBS and only in 4.8% of equine AD-MSC cultivated with 10% ePL. Clonal aberrations in the AD-MSC were neither observed in FBS nor in 10% ePL medium. Analysis of AD-MSC and endothelial cells in an indirect co-culture revealed that the ePL supported the pro-angiogenic effects of AD-MSC. In the 10% ePL group, more vascular endothelial growth factor (VEGF-A) was released and highest VEGF-A concentrations were reached in the presence of ePL and co-cultured cells (p < 0.05). Correspondingly, AD-MSC expressed the VEGF receptor-2 at higher levels in the presence of ePL (p < 0.05). Finally, AD-MSC and 10% ePL together promoted the growth of endothelial cells and induced the formation of vessel-like structures in two of the samples. These data further substantiate that buffy-coat-based ePL is a valuable supplement for equine AD-MSC culture media. The ePL does not only support stable equine AD-MSC characteristics as demonstrated before, but it also enhances their functional properties. |
format | Online Article Text |
id | pubmed-9395693 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-93956932022-08-24 Functional properties of equine adipose-derived mesenchymal stromal cells cultured with equine platelet lysate Hagen, Alina Niebert, Sabine Brandt, Vivian-Pascal Holland, Heidrun Melzer, Michaela Wehrend, Axel Burk, Janina Front Vet Sci Veterinary Science Successful translation of multipotent mesenchymal stromal cell (MSC)-based therapies into clinical reality relies on adequate cell production procedures. These should be available not only for human MSC, but also for MSC from animal species relevant to preclinical research and veterinary medicine. The cell culture medium supplementation is one of the critical aspects in MSC production. Therefore, we previously established a scalable protocol for the production of buffy-coat based equine platelet lysate (ePL). This ePL proved to be a suitable alternative to fetal bovine serum (FBS) for equine adipose-derived (AD-) MSC culture so far, as it supported AD-MSC proliferation and basic characteristics. The aim of the current study was to further analyze the functional properties of equine AD-MSC cultured with the same ePL, focusing on cell fitness, genetic stability and pro-angiogenic potency. All experiments were performed with AD-MSC from n = 5 horses, which were cultured either in medium supplemented with 10% FBS, 10% ePL or 2.5% ePL. AD-MSC cultured with 2.5% ePL, which previously showed decreased proliferation potential, displayed higher apoptosis but lower senescence levels as compared to 10% ePL medium (p < 0.05). Non-clonal chromosomal aberrations occurred in 8% of equine AD-MSC cultivated with FBS and only in 4.8% of equine AD-MSC cultivated with 10% ePL. Clonal aberrations in the AD-MSC were neither observed in FBS nor in 10% ePL medium. Analysis of AD-MSC and endothelial cells in an indirect co-culture revealed that the ePL supported the pro-angiogenic effects of AD-MSC. In the 10% ePL group, more vascular endothelial growth factor (VEGF-A) was released and highest VEGF-A concentrations were reached in the presence of ePL and co-cultured cells (p < 0.05). Correspondingly, AD-MSC expressed the VEGF receptor-2 at higher levels in the presence of ePL (p < 0.05). Finally, AD-MSC and 10% ePL together promoted the growth of endothelial cells and induced the formation of vessel-like structures in two of the samples. These data further substantiate that buffy-coat-based ePL is a valuable supplement for equine AD-MSC culture media. The ePL does not only support stable equine AD-MSC characteristics as demonstrated before, but it also enhances their functional properties. Frontiers Media S.A. 2022-08-09 /pmc/articles/PMC9395693/ /pubmed/36016806 http://dx.doi.org/10.3389/fvets.2022.890302 Text en Copyright © 2022 Hagen, Niebert, Brandt, Holland, Melzer, Wehrend and Burk. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Veterinary Science Hagen, Alina Niebert, Sabine Brandt, Vivian-Pascal Holland, Heidrun Melzer, Michaela Wehrend, Axel Burk, Janina Functional properties of equine adipose-derived mesenchymal stromal cells cultured with equine platelet lysate |
title | Functional properties of equine adipose-derived mesenchymal stromal cells cultured with equine platelet lysate |
title_full | Functional properties of equine adipose-derived mesenchymal stromal cells cultured with equine platelet lysate |
title_fullStr | Functional properties of equine adipose-derived mesenchymal stromal cells cultured with equine platelet lysate |
title_full_unstemmed | Functional properties of equine adipose-derived mesenchymal stromal cells cultured with equine platelet lysate |
title_short | Functional properties of equine adipose-derived mesenchymal stromal cells cultured with equine platelet lysate |
title_sort | functional properties of equine adipose-derived mesenchymal stromal cells cultured with equine platelet lysate |
topic | Veterinary Science |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9395693/ https://www.ncbi.nlm.nih.gov/pubmed/36016806 http://dx.doi.org/10.3389/fvets.2022.890302 |
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