Cargando…
Short-term physiological hypoxia potentiates the therapeutic function of mesenchymal stem cells
BACKGROUND: In the bone marrow, MSCs reside in a hypoxic milieu (1–5% O(2)) that is thought to preserve their multipotent state. Typically, in vitro expansion of MSCs is performed under normoxia (~ 21% O(2)), a process that has been shown to impair their function. Here, we evaluated the characterist...
Autores principales: | , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2018
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6180371/ https://www.ncbi.nlm.nih.gov/pubmed/30305185 http://dx.doi.org/10.1186/s13287-018-1007-x |
_version_ | 1783362186747314176 |
---|---|
author | Antebi, Ben Rodriguez, Luis A Walker, Kerfoot P Asher, Amber M Kamucheka, Robin M Alvarado, Lucero Mohammadipoor, Arezoo Cancio, Leopoldo C |
author_facet | Antebi, Ben Rodriguez, Luis A Walker, Kerfoot P Asher, Amber M Kamucheka, Robin M Alvarado, Lucero Mohammadipoor, Arezoo Cancio, Leopoldo C |
author_sort | Antebi, Ben |
collection | PubMed |
description | BACKGROUND: In the bone marrow, MSCs reside in a hypoxic milieu (1–5% O(2)) that is thought to preserve their multipotent state. Typically, in vitro expansion of MSCs is performed under normoxia (~ 21% O(2)), a process that has been shown to impair their function. Here, we evaluated the characteristics and function of MSCs cultured under hypoxia and hypothesized that, when compared to normoxia, dedicated hypoxia will augment the functional characteristics of MSCs. METHODS: Human and porcine bone marrow MSCs were obtained from fresh mononuclear cells. The first study evaluated MSC function following both long-term (10 days) and short-term (48 h) hypoxia (1% O(2)) culture. In our second study, we evaluated the functional characteristics of MSC cultured under short-term 2% and 5% hypoxia. MSCs were evaluated for their metabolic activity, proliferation, viability, clonogenicity, gene expression, and secretory capacity. RESULTS: In long-term culture, common MSC surface marker expression (CD44 and CD105) dropped under hypoxia. Additionally, in long-term culture, MSCs proliferated significantly slower and provided lower yields under hypoxia. Conversely, in short-term culture, MSCs proliferated significantly faster under hypoxia. In both long-term and short-term cultures, MSC metabolic activity was significantly higher under hypoxia. Furthermore, MSCs cultured under hypoxia had upregulated expression of VEGF with concomitant downregulation of HMGB1 and the apoptotic genes BCL-2 and CASP3. Finally, in both hypoxia cultures, the pro-inflammatory cytokine, IL-8, was suppressed, while levels of the anti-inflammatories, IL-1ra and GM-CSF, were elevated in short-term hypoxia only. CONCLUSIONS: In this study, we demonstrate that hypoxia augments the therapeutic characteristics of both porcine and human MSCs. Yet, short-term 2% hypoxia offers the greatest benefit overall, exemplified by the increase in proliferation, self-renewing capacity, and modulation of key genes and the inflammatory milieu as compared to normoxia. These data are important for generating robust MSCs with augmented function for clinical applications. |
format | Online Article Text |
id | pubmed-6180371 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-61803712018-10-18 Short-term physiological hypoxia potentiates the therapeutic function of mesenchymal stem cells Antebi, Ben Rodriguez, Luis A Walker, Kerfoot P Asher, Amber M Kamucheka, Robin M Alvarado, Lucero Mohammadipoor, Arezoo Cancio, Leopoldo C Stem Cell Res Ther Research BACKGROUND: In the bone marrow, MSCs reside in a hypoxic milieu (1–5% O(2)) that is thought to preserve their multipotent state. Typically, in vitro expansion of MSCs is performed under normoxia (~ 21% O(2)), a process that has been shown to impair their function. Here, we evaluated the characteristics and function of MSCs cultured under hypoxia and hypothesized that, when compared to normoxia, dedicated hypoxia will augment the functional characteristics of MSCs. METHODS: Human and porcine bone marrow MSCs were obtained from fresh mononuclear cells. The first study evaluated MSC function following both long-term (10 days) and short-term (48 h) hypoxia (1% O(2)) culture. In our second study, we evaluated the functional characteristics of MSC cultured under short-term 2% and 5% hypoxia. MSCs were evaluated for their metabolic activity, proliferation, viability, clonogenicity, gene expression, and secretory capacity. RESULTS: In long-term culture, common MSC surface marker expression (CD44 and CD105) dropped under hypoxia. Additionally, in long-term culture, MSCs proliferated significantly slower and provided lower yields under hypoxia. Conversely, in short-term culture, MSCs proliferated significantly faster under hypoxia. In both long-term and short-term cultures, MSC metabolic activity was significantly higher under hypoxia. Furthermore, MSCs cultured under hypoxia had upregulated expression of VEGF with concomitant downregulation of HMGB1 and the apoptotic genes BCL-2 and CASP3. Finally, in both hypoxia cultures, the pro-inflammatory cytokine, IL-8, was suppressed, while levels of the anti-inflammatories, IL-1ra and GM-CSF, were elevated in short-term hypoxia only. CONCLUSIONS: In this study, we demonstrate that hypoxia augments the therapeutic characteristics of both porcine and human MSCs. Yet, short-term 2% hypoxia offers the greatest benefit overall, exemplified by the increase in proliferation, self-renewing capacity, and modulation of key genes and the inflammatory milieu as compared to normoxia. These data are important for generating robust MSCs with augmented function for clinical applications. BioMed Central 2018-10-11 /pmc/articles/PMC6180371/ /pubmed/30305185 http://dx.doi.org/10.1186/s13287-018-1007-x Text en © The Author(s). 2018 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 Antebi, Ben Rodriguez, Luis A Walker, Kerfoot P Asher, Amber M Kamucheka, Robin M Alvarado, Lucero Mohammadipoor, Arezoo Cancio, Leopoldo C Short-term physiological hypoxia potentiates the therapeutic function of mesenchymal stem cells |
title | Short-term physiological hypoxia potentiates the therapeutic function of mesenchymal stem cells |
title_full | Short-term physiological hypoxia potentiates the therapeutic function of mesenchymal stem cells |
title_fullStr | Short-term physiological hypoxia potentiates the therapeutic function of mesenchymal stem cells |
title_full_unstemmed | Short-term physiological hypoxia potentiates the therapeutic function of mesenchymal stem cells |
title_short | Short-term physiological hypoxia potentiates the therapeutic function of mesenchymal stem cells |
title_sort | short-term physiological hypoxia potentiates the therapeutic function of mesenchymal stem cells |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6180371/ https://www.ncbi.nlm.nih.gov/pubmed/30305185 http://dx.doi.org/10.1186/s13287-018-1007-x |
work_keys_str_mv | AT antebiben shorttermphysiologicalhypoxiapotentiatesthetherapeuticfunctionofmesenchymalstemcells AT rodriguezluisa shorttermphysiologicalhypoxiapotentiatesthetherapeuticfunctionofmesenchymalstemcells AT walkerkerfootp shorttermphysiologicalhypoxiapotentiatesthetherapeuticfunctionofmesenchymalstemcells AT asheramberm shorttermphysiologicalhypoxiapotentiatesthetherapeuticfunctionofmesenchymalstemcells AT kamuchekarobinm shorttermphysiologicalhypoxiapotentiatesthetherapeuticfunctionofmesenchymalstemcells AT alvaradolucero shorttermphysiologicalhypoxiapotentiatesthetherapeuticfunctionofmesenchymalstemcells AT mohammadipoorarezoo shorttermphysiologicalhypoxiapotentiatesthetherapeuticfunctionofmesenchymalstemcells AT cancioleopoldoc shorttermphysiologicalhypoxiapotentiatesthetherapeuticfunctionofmesenchymalstemcells |