Short-term physiological hypoxia potentiates the therapeutic function of mesenchymal stem cells
Abstract Background In the bone marrow, MSCs reside in a hypoxic milieu (1–5% O2) that is thought to preserve their multipotent state. Typically, in vitro expansion of MSCs is performed under normoxia (~ 21% O2), a process that has been shown to impair their function. Here, we evaluated the characte...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2018-10-01
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| Series: | Stem Cell Research & Therapy |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s13287-018-1007-x |
| _version_ | 1828865762284011520 |
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| author | Ben Antebi Luis A Rodriguez Kerfoot P Walker Amber M Asher Robin M Kamucheka Lucero Alvarado Arezoo Mohammadipoor Leopoldo C Cancio |
| author_facet | Ben Antebi Luis A Rodriguez Kerfoot P Walker Amber M Asher Robin M Kamucheka Lucero Alvarado Arezoo Mohammadipoor Leopoldo C Cancio |
| author_sort | Ben Antebi |
| collection | DOAJ |
| description | Abstract Background In the bone marrow, MSCs reside in a hypoxic milieu (1–5% O2) that is thought to preserve their multipotent state. Typically, in vitro expansion of MSCs is performed under normoxia (~ 21% O2), 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% O2) 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. |
| first_indexed | 2024-12-13T04:35:45Z |
| format | Article |
| id | doaj.art-9a1b051823894bcd9a7c94daf10f3842 |
| institution | Directory Open Access Journal |
| issn | 1757-6512 |
| language | English |
| last_indexed | 2024-12-13T04:35:45Z |
| publishDate | 2018-10-01 |
| publisher | BMC |
| record_format | Article |
| series | Stem Cell Research & Therapy |
| spelling | doaj.art-9a1b051823894bcd9a7c94daf10f38422022-12-21T23:59:27ZengBMCStem Cell Research & Therapy1757-65122018-10-019111510.1186/s13287-018-1007-xShort-term physiological hypoxia potentiates the therapeutic function of mesenchymal stem cellsBen Antebi0Luis A Rodriguez1Kerfoot P Walker2Amber M Asher3Robin M Kamucheka4Lucero Alvarado5Arezoo Mohammadipoor6Leopoldo C Cancio7United States Army Institute of Surgical ResearchUnited States Army Institute of Surgical ResearchUnited States Army Institute of Surgical ResearchUnited States Army Institute of Surgical ResearchUnited States Army Institute of Surgical ResearchUnited States Army Institute of Surgical ResearchUnited States Army Institute of Surgical ResearchUnited States Army Institute of Surgical ResearchAbstract Background In the bone marrow, MSCs reside in a hypoxic milieu (1–5% O2) that is thought to preserve their multipotent state. Typically, in vitro expansion of MSCs is performed under normoxia (~ 21% O2), 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% O2) 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.http://link.springer.com/article/10.1186/s13287-018-1007-xMesenchymal stem cells (MSCs)HypoxiaNormoxiaBone marrowImmunomodulation |
| spellingShingle | Ben Antebi Luis A Rodriguez Kerfoot P Walker Amber M Asher Robin M Kamucheka Lucero Alvarado Arezoo Mohammadipoor Leopoldo C Cancio Short-term physiological hypoxia potentiates the therapeutic function of mesenchymal stem cells Stem Cell Research & Therapy Mesenchymal stem cells (MSCs) Hypoxia Normoxia Bone marrow Immunomodulation |
| 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 | Mesenchymal stem cells (MSCs) Hypoxia Normoxia Bone marrow Immunomodulation |
| url | http://link.springer.com/article/10.1186/s13287-018-1007-x |
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