In vitro aging alters the gene expression and secretome composition of canine adipose-derived mesenchymal stem cells
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PROVIDER: PRJNA1075668 | ENA |
REPOSITORIES: ENA
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Project description:Canine adipose-derived mesenchymal stem cells (cAD-MSCs) show therapeutic promise for their regenerative potential, particularly in their secretome. However, concerns arise regarding the impact of in vitro cultivation need for storing therapeutic doses, prompting this study to comprehensively explore the impact of in vitro aging on gene expression and secretome composition. The study involved collecting abdominal adipose tissue samples from nine healthy female dogs, from which cAD-MSCs were extracted and cultured. Stem cells were validated through trilineage differentiation assays and flow cytometry immunophenotyping. Gene expression profiling, using RT-qPCR array and cAD-MSCs secretome LC-MS/MS analysis, were conducted at passages 3 and 6 to reveal gene expression and protein composition alterations during in vitro culture. Gene expression profiling of in vitro aged cAD-MSCs revealed expression alterations, while significant downregulation was observed in two MSC-associated genes. Proteomic analysis revealed 10% distinctively expressed proteins, and several up- and downregulations. Grouping these proteins with biologically significant ones, Gene Ontology Panther Pathway analysis revealed that P3 proteins were significantly associated with cytoskeletal regulation, nicotinic acetylcholine receptor signaling, inflammation mediated by chemokine and cytokine signaling, Wnt signaling, and CCKR signaling map. In contrast, P6 proteins were linked to the xanthine and guanine salvage pathway, adenine and hypoxanthine salvage pathway, and blood coagulation pathway. To the best of our knowledge, this study presents the first original perspective on the changes in secretome composition that occur when cAD-MSCs age in vitro. Our findings highlight significant changes that, in conclusion, indicate the regenerative potential of cAD-MSCs and that their secretome may be compromised due to in vitro aging. Consequently, our study suggests a preference for earlier passages when considering these cells for therapeutic applications.
2024-02-15 | GSE255585 | GEO
Project description:Canine adipose-derived mesenchymal stem cells (cAD-MSCs) show therapeutic promise for their regenerative potential, particularly in their secretome. However, concerns arise regarding the impact of in vitro cultivation need for storing therapeutic doses, prompting this study to comprehensively explore the impact of in vitro aging on gene expression and secretome composition. The study involved collecting abdominal adipose tissue samples from nine healthy female dogs, from which cAD-MSCs were extracted and cultured. Stem cells were validated through trilineage differentiation assays and flow cytometry immunophenotyping. Gene expression profiling, using RT-qPCR array and cAD-MSCs secretome LC-MS/MS analysis, were conducted at passages 3 and 6 to reveal gene expression and protein composition alterations during in vitro culture. Gene expression profiling of in vitro aged cAD-MSCs revealed expression alterations, while significant downregulation was observed in two MSC-associated genes. Proteomic analysis revealed 10% distinctively expressed proteins, and several up- and downregulations. Grouping these proteins with biologically significant ones, Gene Ontology Panther Pathway analysis revealed that P3 proteins were significantly associated with cytoskeletal regulation, nicotinic acetylcholine receptor signaling, inflammation mediated by chemokine and cytokine signaling, Wnt signaling, and CCKR signaling map. In contrast, P6 proteins were linked to the xanthine and guanine salvage pathway, adenine and hypoxanthine salvage pathway, and blood coagulation pathway. To the best of our knowledge, this study presents the first original perspective on the changes in secretome composition that occur when cAD-MSCs age in vitro. Our findings highlight significant changes that, in conclusion, indicate the regenerative potential of cAD-MSCs and that their secretome may be compromised due to in vitro aging. Consequently, our study suggests a preference for earlier passages when considering these cells for therapeutic applications.
2024-03-13 | PXD049324 | Pride
Project description:IntroductionCanine adipose-derived mesenchymal stem cells (cAD-MSCs) hold therapeutic promise due to their regenerative potential, particularly within their secretome. However, concerns arise regarding the impact of in vitro cultivation necessitated for storing therapeutic doses, prompting this study to comprehensively explore the impact of in vitro aging on gene expression and secretome composition.MethodsThe study involved collecting abdominal adipose tissue samples from nine healthy female dogs, from which cAD-MSCs were extracted and cultured. Stem cells were validated through trilineage differentiation assays and flow cytometry immunophenotyping. Gene expression profiling using RT-qPCR array, and cAD-MSCs secretome LC-MS/MS analysis, were conducted at passages 3 and 6 to reveal gene expression and protein composition alterations during in vitro culture.Results and discussionThe results demonstrate that the gene expression and secretome composition of cAD-MSCs were impacted by in vitro aging. Among many alterations in gene expression between two passages, two significant downregulations were noted in the MSC-associated PTPRC and IL10 genes. While the majority of proteins and their functional characteristics were shared between passages, the influence of cell aging on secretome composition is highlighted by 10% of proteins being distinctively expressed in each passage, along with 21 significant up- and downregulations. The functional attributes of proteins detected in passage 3 demonstrated a greater inclination towards supporting the regenerative capacity of cAD-MSCs. Moreover, proteins in passage 6 exhibited a noteworthy correlation with the blood coagulation pathway, suggesting an elevated likelihood of coagulation events. To the best of our knowledge, this study presents the first original perspective on the changes in secretome composition that occur when cAD-MSCs age in vitro. Furthermore, it contributes to broadening the currently restricted knowledge base concerning the secretome of cAD-MSCs. In conclusion, our findings show that the regenerative potential of cAD-MSCs, as well as their secretome, may be compromised by in vitro aging. Therefore, our study suggests a preference for earlier passages when considering these cells for therapeutic applications.
| S-EPMC11006985 | biostudies-literature
Project description:Canine adipose -derived mesenchymal stem cells (cAD-MSCs) demonstrate promising tissue repair and regeneration capabilities. However, the procurement and preservation of these cells or their secreted factors for therapeutic applications pose a risk of viral contamination, and the consequences on cAD-MSCs remain unexplored. Consequently, this research sought to assess the impact of Canid alphaherpesvirus 1 on the functional attributes of cAD-MSCs, including gene expression profiles and secretome composition. To this end, abdominal fat tissue from twelve healthy dogs was harvested to isolate cAD-MSCs. These samples were tested for CHV contamination before introducing a wild-type CHV strain via serial passages. Following CHV infection, RT-PCR arrays and LC-MS/MS assessments enabled gene expression analysis and the secretome's proteomic landscape, respectively. The study established that the initial cAD-MSCs populations were devoid of CHV. Upon exposure to the virus, cAD-MSCs exhibited susceptibility, leading to notable modifications in gene expression and secretome profile. These changes reflected their intrinsic properties, such as stemness, differentiation potential, structural integrity, proliferative capacity, survival, directional migration, and immune-modulatory functions, depriving their regenerative efficacy . Despite the absence of pre-existing contamination, the outcomes underscore the imperative of routine viral screening prior to therapeutic use, particularly CHV. Moreover, these findings provide insights into the pathogenic mechanisms of CHV and the intricate interactions between canine stem cells and viruses.
2024-05-17 | GSE267402 | GEO
Project description:The adult stem cells have become prominent candidates for the treatment of various diseases in veterinary practice. Therefore, the main goal of our study was to provide a complex study of canine bone marrow derived mesenchymal stem cells (BMMSCs) and conditioned media, isolated from healthy adult dogs of different breeds. Under well-defined standardized isolation protocols the multilineage differentiation and specific surface markers of MSCs were supplemented with their gene expression and the proteome profile. Present data has confirmed that canine BMMSCs expresses important genes for differentiation toward osteo-, chondro- and tendo-genic directions, but also genes that were associated with neurotrophic and immunomodulatory properties. Furthermore, we have identified for the first time by proteome profiling dynamic release various bioactive molecules, such as transcription and translation factors, osteogenic, growth and neurotrophic factors from canine BMMSC conditioned medium. Importantly, relevant genes were linked to their proteins that were detected in conditioned medium, which have an important correlation to their possible function and clinical application. Thus, here we show that the canine BMMSCs properties are applicable for replacement therapies of skeletal disease, but due to their ability of releasing immunomodulatory and bioactive molecules via conditioned media, they may improve repair of other tissue. However, extensive experimental or pre-clinical trials testing canine sources needs to be performed in order to better understand the individual differences between healthy donors that are considered for transplantation treatment studies in veterinary medicine.
2019-07-15 | PXD013058 | Pride
Project description:Canine adipose -derived mesenchymal stem cells (cAD-MSCs) demonstrate promising tissue repair and regeneration capabilities. However, the procurement and preservation of these cells or their secreted factors for therapeutic applications pose a risk of viral contamination, and the consequences on cAD-MSCs remain unexplored. Consequently, this research sought to assess the impact of Canid alphaherpesvirus 1 on the functional attributes of cAD-MSCs, including gene expression profiles and secretome composition. To this end, abdominal fat tissue from twelve healthy dogs was harvested to isolate cAD-MSCs. These samples were tested for CHV contamination before introducing a wild-type CHV strain via serial passages. Following CHV infection, RT-PCR arrays and LC-MS/MS assessments enabled gene expression analysis and the secretome's proteomic landscape, respectively. The study established that the initial cAD-MSCs populations were devoid of CHV. Upon exposure to the virus, cAD-MSCs exhibited susceptibility, leading to notable modifications in gene expression and secretome profile. These changes reflected their intrinsic properties, such as stemness, differentiation potential, structural integrity, proliferative capacity, survival, directional migration, and immune-modulatory functions, depriving their regenerative efficacy . Despite the absence of pre-existing contamination, the outcomes underscore the imperative of routine viral screening prior to therapeutic use, particularly CHV. Moreover, these findings provide insights into the pathogenic mechanisms of CHV and the intricate interactions between canine stem cells and viruses.
2024-12-16 | PXD052289 | Pride
Project description:Mesenchymal stromal cells (MSCs) constitute the cell type more frequently used in many regenerative medicine approaches due to their exclusive immunomodulatory properties, and they have been reported to mediate profound immunomodulatory effects in vivo. Nevertheless, MSCs do not express essential adhesion molecules actively involved in cell migration, a phenotypic feature that hampers their ability to home inflamed tissues following intravenous administration. In this study, we investigated whether modification by fucosylation of murine AdMSCs (mAdMSCs) creates Hematopoietic Cell E-/L-selectin Ligand, the E-selectin-binding CD44 glycoform. This cell surface glycan modification of CD44 has previously shown in preclinical studies to favor trafficking of mAdMSCs to inflamed or injured peripheral tissues. We analyzed the impact that exofucosylation could have in other innate phenotypic and functional properties of MSCs. Compared to unmodified counterparts, fucosylated mAdMSCs demonstrated higher in vitro migration, an altered secretome pattern, including increased expression and secretion of anti-inflammatory molecules, and a higher capacity to inhibit mitogen-stimulated splenocyte proliferation under standard culture conditions. Together, these findings indicate that exofucosylation could represent a suitable cell engineering strategy, not only to facilitate the in vivo MSC colonization of damaged tissues after systemic administration, but also to convert MSCs in a more potent immunomodulatory/anti-inflammatory cell therapy-based product for the treatment of a variety of autoimmune, inflammatory, and degenerative diseases.
| S-EPMC7726227 | biostudies-literature
Project description:BackgroundCanine adipose-derived mesenchymal stem cells (cAD-MSCs) demonstrate promising tissue repair and regeneration capabilities. However, the procurement and preservation of these cells or their secreted factors for therapeutic applications pose a risk of viral contamination, and the consequences for cAD-MSCs remain unexplored. Consequently, this research sought to assess the impact of canid alphaherpesvirus 1 (CHV) on the functional attributes of cAD-MSCs, including gene expression profiles and secretome composition.MethodsTo this end, abdominal adipose tissue from 12 healthy dogs was harvested to isolate cAD-MSCs. These samples were tested for CHV contamination before introducing a wild-type CHV strain via serial passages. Following CHV infection, real-time reverse transcription-polymerase chain reaction array and liquid chromatography with tandem mass spectrometry assessments enabled analyses of gene expression and secretome's proteomic profile, respectively.ResultsThis study showed that the initial cAD-MSC populations were devoid of CHV. cAD-MSCs showed susceptibility to infection with wild-type CHV, leading to notable modifications in gene expression and secretome profile. The observed genomic variations in gene expression indicate potential impacts on the stemness, migration, and other functional properties of cAD-MSCs, highlighting the need for further studies to evaluate their functional capacity post-infection. Moreover, gene expression and secretome analyses suggest a shift in stem cell differentiation toward an adipogenic phenotype.ConclusionTo the best of our knowledge, this is the first study of the effects of virus infection on gene expression and secretome composition in cAD-MSCs. The outcomes of our study underscore the imperative of routine viral screening prior to the therapeutic use of cAD-MSCs. Moreover, these findings provide novel insights into the pathogenic mechanisms of CHV and pave the way for future canine stem cell and virus research.
| S-EPMC11673362 | biostudies-literature
Project description:Osteoarthritis (OA) is a degenerative joint disease characterized by cartilage degradation and inflammation. This study investigates the therapeutic potential of secretome derived from adipose tissue mesenchymal stem cells (ASCs) in mitigating inflammation and promoting cartilage repair in an in vitro model of OA. Our in vitro model comprised chondrocytes inflamed with TNF. To assess the therapeutic potential of secretome, inflamed chondrocytes were treated with it and concentrations of pro-inflammatory cytokines, metalloproteinases (MMPs) and extracellular matrix markers were measured. In addition, secretome-treated chondrocytes were subject to a microarray analysis to determine which genes were upregulated and which were downregulated. Treating TNF-inflamed chondrocytes with secretome in vitro inhibits the NF-κB pathway, thereby mediating anti-inflammatory and anti-catabolic effects. Additional protective effects of secretome on cartilage are revealed in the inhibition of hypertrophy markers such as RUNX2 and COL10A1, increased production of COL2A1 and ACAN and upregulation of SOX9. These findings suggest that ASC-derived secretome can effectively reduce inflammation, promote cartilage repair, and maintain chondrocyte phenotype. This study highlights the potential of ASC-derived secretome as a novel, non-cell-based therapeutic approach for OA, offering a promising alternative to current treatments by targeting inflammation and cartilage repair mechanisms.
2024-10-22 | GSE239343 | GEO
Project description:The objective of this project is secreening the constituent of adipose derived mesenchymal stem cell secretome.
2023-09-11 | PXD045254 |