In vitro aging alters the gene expression and secretome composition of canine adipose-derived mesenchymal stem cells
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA1075668 | ENA |
REPOSITORIES: ENA
Similar Datasets
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: Not available
| 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:Mesenchymal Stromal Cells (MSCs) are widely studied in regenerative medicine because of their unique biological properties. The MSC-derived secretome is a cell-free product containing soluble factors and extracellular vesicles that mediate many of the biological properties of MSCs, including immunomodulatory and antimicrobial activities. This study aimed to evaluate the antimicrobial activity of a freeze-dried secretome derived from canine adipose tissue MSCs (canine lyosecretome, c-Lyo) against pathogens relevant to canine infections. The antimicrobial activity of c-Lyo was assessed using minimal inhibitory concentration (MIC) assays against six bacterial species (Escherichia coli, Salmonella enterica subsp. enterica serovar Typhimurium, Staphylococcus aureus, methicillin-resistant Staphylococcus aureus (MRSA), Staphylococcus pseudintermedius and Pseudomonas aeruginosa) and one yeast (Malassezia pachydermatis). Potential interactions between c-Lyo and commonly used veterinary antimicrobials (gentamicin, amoxicillin, enrofloxacin, and ketoconazole) were evaluated using checkerboard assays (CkA). Proteomic characterization of the c-Lyo and transcriptomic analysis of canine MSCs were also conducted to explore molecular mechanisms underlying antimicrobial activity. The results demonstrated that c-Lyo exerts antimicrobial activity against all tested microorganisms, with variable sensitivity among species. CkA exhibited predominantly additive interactions with conventional antimicrobials and occasional synergistic effects, with no evidence of antagonism. Proteomic and transcriptomic analyses supported the presence of antimicrobial peptides and immune-related pathways that potentially contribute to both direct antimicrobial activity and the modulation of host defense mechanisms. These findings indicate that c-Lyo may represent a promising cell-free biologic approach for antimicrobial strategies in veterinary medicine.
2026-04-09 | GSE327506 | 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: Not available
| S-EPMC7726227 | biostudies-literature
Project description: Not available
| 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