Functional properties of equine adipose-derived mesenchymal stromal cells cultured with equine platelet lysate.
Abstract: Successful translation of multipotent mesenchymal stromal cell (MSC)-based therapies into clinical reality relies on adequate cell production procedures. These should be available not only for human MSC, but also for MSC from animal species relevant to preclinical research and veterinary medicine. The cell culture medium supplementation is one of the critical aspects in MSC production. Therefore, we previously established a scalable protocol for the production of buffy-coat based equine platelet lysate (ePL). This ePL proved to be a suitable alternative to fetal bovine serum (FBS) for equine adipose-derived (AD-) MSC culture so far, as it supported AD-MSC proliferation and basic characteristics. The aim of the current study was to further analyze the functional properties of equine AD-MSC cultured with the same ePL, focusing on cell fitness, genetic stability and pro-angiogenic potency. All experiments were performed with AD-MSC from = 5 horses, which were cultured either in medium supplemented with 10% FBS, 10% ePL or 2.5% ePL. AD-MSC cultured with 2.5% ePL, which previously showed decreased proliferation potential, displayed higher apoptosis but lower senescence levels as compared to 10% ePL medium ( < 0.05). Non-clonal chromosomal aberrations occurred in 8% of equine AD-MSC cultivated with FBS and only in 4.8% of equine AD-MSC cultivated with 10% ePL. Clonal aberrations in the AD-MSC were neither observed in FBS nor in 10% ePL medium. Analysis of AD-MSC and endothelial cells in an indirect co-culture revealed that the ePL supported the pro-angiogenic effects of AD-MSC. In the 10% ePL group, more vascular endothelial growth factor (VEGF-A) was released and highest VEGF-A concentrations were reached in the presence of ePL and co-cultured cells ( < 0.05). Correspondingly, AD-MSC expressed the VEGF receptor-2 at higher levels in the presence of ePL ( < 0.05). Finally, AD-MSC and 10% ePL together promoted the growth of endothelial cells and induced the formation of vessel-like structures in two of the samples. These data further substantiate that buffy-coat-based ePL is a valuable supplement for equine AD-MSC culture media. The ePL does not only support stable equine AD-MSC characteristics as demonstrated before, but it also enhances their functional properties.
Copyright © 2022 Hagen, Niebert, Brandt, Holland, Melzer, Wehrend and Burk.
Publication Date: 2022-08-09 PubMed ID: 36016806PubMed Central: PMC9395693DOI: 10.3389/fvets.2022.890302Google Scholar: Lookup
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Summary
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This study investigated the use of equine platelet lysate (ePL) in culturing equine adipose-derived mesenchymal stromal cells (AD-MSCs), exploring the effects of ePL on cell behavior, genetic stability, and angiogenic potential. It concludes that ePL not only maintains the stable characteristics of equine AD-MSCs but also enhances their functional properties.
Cell Culturing and Adipose-Derived Mesenchymal Stromal Cells
- The research highlights the importance of effective cell production procedures in translating the potential of mesenchymal stromal cell (MSC) therapies into clinical reality. Such procedures need to be developed and tailored not just for human MSCs, but also for those from animal species in areas like veterinary medicine and preclinical research.
- The study focuses on equine adipose-derived mesenchymal stromal cells (AD-MSCs). These are cells isolated from the adipose (fat) tissue of horses, which have certain abilities like differentiation and self-renewal, making them a focus of many therapeutic and scientific research.
Role of Equine Platelet Lysate in Cell Culture
- Equine platelet lysate (ePL) is an alternative to fetal bovine serum (FBS), a supplement often used in cell culture. The ePL was previously found to support the proliferation and basic characteristics of equine AD-MSCs.
- In this study, the effects of ePL on the functional properties of AD-MSCs were further analyzed, with a focus on cell fitness, genetic stability, and pro-angiogenic potency – i.e., the capacity to promote new blood vessel formation.
Findings and Conclusions
- AD-MSCs cultured with 2.5% ePL showed increased rates of apoptosis (programmed cell death) and decreased senescence levels compared to those cultured with 10% ePL.
- Non-clonal chromosomal aberrations (random changes in chromosome structure or number) occurred less frequently in AD-MSCs cultured with ePL than in those cultured with FBS, suggesting that ePL might promote greater genetic stability.
- ePL was found to support the pro-angiogenic effects of the AD-MSCs. In the presence of ePL, the cells exhibited increased expression of the vascular endothelial growth factor (VEGF) receptor-2, a key protein involved in angiogenesis. Furthermore, the ePL and the AD-MSCs together promoted the growth of endothelial cells and led to the formation of vessel-like structures in some samples.
- These findings suggest that ePL is not only a feasible substitute for FBS in equine AD-MSC culture media, but it could actually enhance the functional properties of the cells.
Cite This Article
APA
Hagen A, Niebert S, Brandt VP, Holland H, Melzer M, Wehrend A, Burk J.
(2022).
Functional properties of equine adipose-derived mesenchymal stromal cells cultured with equine platelet lysate.
Front Vet Sci, 9, 890302.
https://doi.org/10.3389/fvets.2022.890302 Publication
Researcher Affiliations
- Equine Clinic (Surgery, Orthopedics), Justus-Liebig-University Giessen, Giessen, Germany.
- Equine Clinic (Surgery, Orthopedics), Justus-Liebig-University Giessen, Giessen, Germany.
- Saxon Incubator for Clinical Translation (SIKT), University of Leipzig, Leipzig, Germany.
- Saxon Incubator for Clinical Translation (SIKT), University of Leipzig, Leipzig, Germany.
- Equine Clinic (Surgery, Orthopedics), Justus-Liebig-University Giessen, Giessen, Germany.
- Clinic for Obstetrics, Gynecology and Andrology of Large and Small Animals, Justus-Liebig-University Giessen, Giessen, Germany.
- Equine Clinic (Surgery, Orthopedics), Justus-Liebig-University Giessen, Giessen, Germany.
Conflict of Interest Statement
The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
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Citations
This article has been cited 3 times.- Moellerberndt J, Hagen A, Niebert S, Büttner K, Burk J. Cytokines in equine platelet lysate and related blood products. Front Vet Sci 2023;10:1117829.
- Duysens J, Graide H, Niesten A, Mouithys-Mickalad A, Deby-Dupont G, Franck T, Ceusters J, Serteyn D. Culture and Immunomodulation of Equine Muscle-Derived Mesenchymal Stromal Cells: A Comparative Study of Innovative 2D versus 3D Models Using Equine Platelet Lysate. Cells 2024 Jul 31;13(15).
- Moellerberndt J, Niebert S, Fey K, Hagen A, Burk J. Impact of platelet lysate on immunoregulatory characteristics of equine mesenchymal stromal cells. Front Vet Sci 2024;11:1385395.
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