Mass spectrometric analysis of the in vitro secretome from equine bone marrow-derived mesenchymal stromal cells to assess the effect of chondrogenic differentiation on response to interleukin-1β treatment.
Abstract: Similar to humans, the horse is a long-lived, athletic species. The use of mesenchymal stromal cells (MSCs) is a relatively new frontier, but has been used with promising results in treating joint diseases, e.g., osteoarthritis. It is believed that MSCs exert their main therapeutic effects through secreted trophic biomolecules. Therefore, it has been increasingly important to characterize the MSC secretome. It has been shown that the effect of the MSCs is strongly influenced by the environment in the host compartment, and it is a crucial issue when considering MSC therapy. The aim of this study was to investigate differences in the in vitro secreted protein profile between naïve and chondrogenic differentiating bone marrow-derived (BM)-MSCs when exposed to an inflammatory environment. Equine BM-MSCs were divided into a naïve group and a chondrogenic group. Cells were treated with normal expansion media or chondrogenic media. Cells were treated with IL-1β for a period of 5 days (stimulation), followed by 5 days without IL-1β (recovery). Media were collected after 48 h and 10 days. The secretomes were digested and analyzed by nanoLC-MS/MS to unravel the orchestration of proteins. The inflammatory proteins IL6, CXCL1, CXCL6, CCL7, SEMA7A, SAA, and haptoglobin were identified in the secretome after 48 h from all cells stimulated with IL-1β. CXCL8, OSM, TGF-β1, the angiogenic proteins VCAM1, ICAM1, VEGFA, and VEGFC, the proteases MMP1 and MMP3, and the protease inhibitor TIMP3 were among the proteins only identified in the secretome after 48 h from cells cultured in normal expansion media. After 10-day incubation, the proteins CXCL1, CXCL6, and CCL7 were still identified in the secretome from BM-MSCs stimulated with IL-1β, but the essential inducer of inflammation, IL6, was only identified in the secretome from cells cultured in normal expansion media. The findings in this study indicate that naïve BM-MSCs have a more extensive inflammatory response at 48 h to stimulation with IL-1β compared to BM-MSCs undergoing chondrogenic differentiation. This extensive inflammatory response decreased after 5 days without IL-1β (day 10), but a difference in composition of the secretome between naïve and chondrogenic BM-MSCs was still evident.
Publication Date: 2020-05-20 PubMed ID: 32434555PubMed Central: PMC7238576DOI: 10.1186/s13287-020-01706-7Google Scholar: Lookup
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- Journal Article
Summary
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This research examines the differences in protein secretion profiles between undifferentiated (naïve) and chondrogenic differentiating bone marrow-derived mesenchymal stromal cells (BM-MSCs) of horses when exposed to an inflammatory environment. The findings suggest that naïve BM-MSCs have a stronger inflammatory response to stimulation with the inflammatory protein IL-1β compared to those undergoing chondrogenic differentiation, which could have implications for MSC therapy.
Objective of the Study
- The aim of this study was to scrutinize difference of the secreted protein profile in the naïve and chondrogenic differentiating BM-MSCs when they are exposed to an inflammatory environment. The bone marrow derived mesenchymal stromal cell (MSC) therapy is a newer stream in medicine and is used to treat joint diseases like osteoarthritis. These MSCs mainly work through secreting therapeutic biomolecules, hence understanding their ‘secretome’ has been seen as important.
Methodology
- Equine BM-MSCs were divided into two groups: a naïve group and a chondrogenic group. The cells were treated with either regular expansion media or chondrogenic media.
- They were then exposed to inflammatory protein IL-1β for a period of 5 days (stimulation phase) and then kept for another 5 days without IL-1β (recovery phase).
- After 48 hours and 10 days, the media were collected and analyzed using nanoLC-MS/MS, a highly sensitive technique for identifying and quantifying proteins.
Results
- The study found a range of inflammatory proteins like IL6, CXCL1, CXCL6, CCL7, SEMA7A, SAA, and haptoglobin in the secretome (the group of proteins secreted by the cells) from all cells treated with IL-1β after 48 hours.
- Proteins like CXCL8, OSM, TGF-β1, VCAM1, ICAM1, VEGFA, VEGFC, MMP1, MMP3, and TIMP3 were only identified in the secretome after 48 hours from cells cultured in normal expansion media.
- Even after 10 days of incubation, CXCL1, CXCL6, and CCL7 proteins were still found in the secretome from MSCs stimulated with IL-1β. IL6, an inducer of inflammation, was only identified in the secretome from cells cultured in normal expansion media.
- The findings suggest that naïve MSCs have a stronger inflammatory response after 48 hours of exposure to IL-1β than those undergoing differentiation. This response decreases after 5 days without IL-1β but a difference can still be observed in the secretome composition of the two groups after 10 days.
Conclusion
- The results of the study provide valuable insights into the differences in protein secretion profiles between the naïve and chondrogenic differentiating BM-MSCs under an inflammatory environment.
- The study adds to the growing body of research on the potential therapeutic applications of MSCs, shedding light on their behavior under inflammatory conditions, like those seen in clinical scenarios such as osteoarthritis.
Cite This Article
APA
Bundgaard L, Stensballe A, Elbæk KJ, Berg LC.
(2020).
Mass spectrometric analysis of the in vitro secretome from equine bone marrow-derived mesenchymal stromal cells to assess the effect of chondrogenic differentiation on response to interleukin-1β treatment.
Stem Cell Res Ther, 11(1), 187.
https://doi.org/10.1186/s13287-020-01706-7 Publication
Researcher Affiliations
- Department of Veterinary Clinical Sciences, University of Copenhagen, Agrovej 8, 2630, Taastrup, Denmark. lb@sund.ku.dk.
- Department of Health Science and Technology, Aalborg University, Fredrik Bajers Vej 7E, 9220, Aalborg Ø, Denmark.
- Department of Health Science and Technology, Aalborg University, Fredrik Bajers Vej 7E, 9220, Aalborg Ø, Denmark.
- Department of Veterinary Clinical Sciences, University of Copenhagen, Agrovej 8, 2630, Taastrup, Denmark.
MeSH Terms
- Animals
- Bone Marrow
- Bone Marrow Cells
- Cell Differentiation
- Cells, Cultured
- Horses
- Interleukin-1beta
- Mesenchymal Stem Cells
- Tandem Mass Spectrometry
Conflict of Interest Statement
The authors declare that they have no competing interests.
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