The Expression Pattern of Surface Markers in Canine Adipose-Derived Mesenchymal Stem Cells

Krešić, Nina; Prišlin, Marina; Vlahović, Dunja; Kostešić, Petar; Ljolje, Ivana; Brnić, Dragan; Turk, Nenad; Musulin, Andrija; Habrun, Boris

doi:10.3390/ijms22147476

Cited by 10 publications

(10 citation statements)

References 41 publications

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“…During the analysis, we found a lower percentage of CD29+, CD44+ and CD105+ and almost no positivity for CD90 in AM-MSCs compared with BM-MSCs and AT-MCs, where the expression of all four CD markers was high [ 35 ]. The majority of canine papers have demonstrated either a single alternative MSC marker (CD29 or CD44) or both, which is more consistent than any of the classic MSC markers [ 15 , 23 , 24 , 36 ]. High expression of CD29 and CD105 is superior to strong chondrogenic potential by regulating TGF-β/Smad2 [ 37 ].…”

Section: Discussionmentioning

confidence: 98%

A Comparative Study of Canine Mesenchymal Stem Cells Isolated from Different Sources

Humeník

Maloveská

Hudáková

et al. 2022

Animals

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In this study, we provide comprehensive analyses of mesenchymal stem cells (MSCs) isolated from three types of canine tissues: bone marrow (BM-MSCs), adipose tissue (AT-MSCs) and amniotic tissue (AM-MSCs). We compare their morphology, phenotype, multilineage potential and proliferation activity. The BM-MSCs and AM-MSCs showed fibroblast-like shapes against the spindle shape of the AT-MSCs. All populations showed strong osteogenic and chondrogenic potential. However, we observed phenotypic differences. The BM-MSCs and AT-MSCs revealed high expression of CD29, CD44, CD90 and CD105 positivity compared to the AM-MSCs, which showed reduced expression of all the analysed CD markers. Similarly, the isolation yield and proliferation varied depending on the source. The highest isolation yield and proliferation were detected in the population of AT-MSCs, while the AM-MSCs showed a high yield of cells, but the lowest proliferation activity, in contrast to the BM-MSCs which had the lowest isolation yield. Thus, the present data provide assumptions for obtaining a homogeneous MSC derived from all three canine tissues for possible applications in veterinary regenerative medicine, while the origin of isolated MSCs must always be taken into account.

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Section: Discussionmentioning

confidence: 98%

A Comparative Study of Canine Mesenchymal Stem Cells Isolated from Different Sources

Humeník

Maloveská

Hudáková

et al. 2022

Animals

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“…Nevertheless, sex was not recognized as influential in cell surface marker expression, viability, proliferation, or differentiation potential ( 21 ). The acquisition of adipose tissue, cAD-MSCs extraction, and propagation adhered to the established protocol previously detailed in Krešić et al ( 18 ). Briefly, 1–7 g of adipose tissue ( Table 1 ), depending on the availability, was obtained from biomedical waste of ovarian mesostructure, placed into a sterile Falcon tube devoid of the medium, and stored at 4°C until transportation to the laboratory.…”

Section: Methodsmentioning

confidence: 99%

“…In previous research ( 18 ), the multipotency of the extracted cAD-MSCs was evaluated through trilineage differentiation, encompassing adipogenic, osteogenic, and chondrogenic lineages. This assessment was conducted in the third passage (P3).…”

Section: Methodsmentioning

confidence: 99%

“…In contrast to human MSCs, there has been limited investigation into the impact of in vitro aging on gene expression profiles and secretome composition of cAD-MSCs. Our prior research has contributed to a better understanding of gene expression changes during in vitro passages, albeit on a limited scale ( 17 , 18 ), indicating the need for further investigation. To our knowledge, no data has been published regarding alterations in the secretome composition during long-term culture of cAD-MSCs.…”

Section: Introductionmentioning

confidence: 99%

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In vitro aging alters the gene expression and secretome composition of canine adipose-derived mesenchymal stem cells

Prišlin,

Butorac,

Bertoša

et al. 2024

Front. Vet. Sci.

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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.

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“…Others have demonstrated a variation in CD73 expression in early and late culture passages of canine BM-MSCs, with a significant decrease of expression in late phases. This indicates that extensive in vitro expansion is a factor conditioning the surface marker profile of MSCs (179), which could be intuitive given the fact that cells are isolated from a complex 3D environment rich with intercellular and paracrine stimuli, to a planar 2D environment resulting in a homogeneous cell population in a controlled environment.…”

Section: Cell Characterizationmentioning

confidence: 96%

Manufacturing Mesenchymal Stromal Cells for the Treatment of Osteoarthritis in Canine Patients: Challenges and Recommendations

et al. 2022

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The recent interest in advanced biologic therapies in veterinary medicine has opened up opportunities for new treatment modalities with considerable clinical potential. Studies with mesenchymal stromal cells (MSCs) from animal species have focused on in vitro characterization (mostly following protocols developed for human application), experimental testing in controlled studies and clinical use in veterinary patients. The ability of MSCs to interact with the inflammatory environment through immunomodulatory and paracrine mechanisms makes them a good candidate for treatment of inflammatory musculoskeletal conditions in canine species. Analysis of existing data shows promising results in the treatment of canine hip dysplasia, osteoarthritis and rupture of the cranial cruciate ligament in both sport and companion animals. Despite the absence of clear regulatory frameworks for veterinary advanced therapy medicinal products, there has been an increase in the number of commercial cell-based products that are available for clinical applications, and currently the commercial use of veterinary MSC products has outpaced basic research on characterization of the cell product. In the absence of quality standards for MSCs for use in canine patients, their safety, clinical efficacy and production standards are uncertain, leading to a risk of poor product consistency. To deliver high-quality MSC products for veterinary use in the future, there are critical issues that need to be addressed. By translating standards and strategies applied in human MSC manufacturing to products for veterinary use, in a collaborative effort between stem cell scientists and veterinary researchers and surgeons, we hope to facilitate the development of quality standards. We point out critical issues that need to be addressed, including a much higher level of attention to cell characterization, manufacturing standards and release criteria. We provide a set of recommendations that will contribute to the standardization of cell manufacturing methods and better quality assurance.

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The Expression Pattern of Surface Markers in Canine Adipose-Derived Mesenchymal Stem Cells

Cited by 10 publications

References 41 publications

A Comparative Study of Canine Mesenchymal Stem Cells Isolated from Different Sources

A Comparative Study of Canine Mesenchymal Stem Cells Isolated from Different Sources

In vitro aging alters the gene expression and secretome composition of canine adipose-derived mesenchymal stem cells

Manufacturing Mesenchymal Stromal Cells for the Treatment of Osteoarthritis in Canine Patients: Challenges and Recommendations

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