Autologous Platelet Lysate Does Not Enhance Chondrogenic Differentiation of Equine Bone Marrow-Derived Mesenchymal Stromal Cells Despite Increased TGF-β1 Concentration

Chapman, Hannah-Sophie; Gale, Alexis L.; Dodson, Michael E.; Linardi, Renata L.; Ortved, Kyla F.

doi:10.1089/scd.2019.0239

Cited by 13 publications

(10 citation statements)

References 46 publications

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“…We could hypothesize that the 3D environment itself provides a larger space for cell growth, and/or the molecular components of serum and platelet concentrates could represent a stimulus for cell replication. As a matter of fact, both 3D fibrin matrix [28] and platelet concentrates [23][24][25][26] represent complex biological cell growth-supplement able to sustain cell survival and replication. Recently, Kakudo et al analyzed the role of PL on human adipose-derived MSCs, demonstrating that the already known proliferative effect is possibly caused by the activation of multiple signaling pathways, as observed for FBS [53].…”

Section: Discussionmentioning

confidence: 99%

“…This approach has been considered in the recent years for the preparation of human MSCs, where the restrictive regulations led either to the formulation of serum-free culture media or to the substitution of FBS by products of human origin, mainly PL [23]. The literature in veterinary medicine, indeed, does not provide much information about the effectiveness of platelet derivatives on MSCs culture and the chance to replace FBS with non-xenobiotic products [24][25][26]. Furthermore, no canine or feline species-specific serum substitutes are commercially available.…”

Section: Introductionmentioning

confidence: 99%

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Xenobiotic-Free Medium Guarantees Expansion of Adipose Tissue-Derived Canine Mesenchymal Stem Cells Both in 3D Fibrin-Based Matrices and in 2D Plastic Surface Cultures

Suelzu

Conti

Khalidy

et al. 2020

Cells

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Mesenchymal stem cells (MSCs) have been recently introduced in veterinary medicine as a potential therapeutic tool for several pathologies. The large-scale in vitro expansion needed to ensure the preparation of a suitable number of MSCs for clinical application usually requires the use of xenogeneic supplements like the fetal bovine serum (FBS). The substitution of FBS with species-specific supplements would improve the safety of implanted cells, reducing the risk of undesired immune responses following cell therapy. We have evaluated the effectiveness of canine adipose tissue-derived stromal vascular fraction (SVF) and MSCs (ADMSCs) expansion in the presence of canine blood-derived supplements. Cells were cultured on traditional plastic surface and inside a 3D environment derived from the jellification of different blood-derived products, i.e., platelet-poor plasma (PPP), platelet-rich plasma (PRP), or platelet lysate (PL). PPP, PRP, and PL can contribute to canine ADMSCs in vitro expansion. Both allogeneic and autologous PPP and PL can replace FBS for ADMSCs culture on a plastic surface, exhibiting either a similar (PPP) or a more effective (PL) stimulus to cell replication. Furthermore, the 3D environment based on homospecific blood-derived products polymerization provides a strong stimulus to ADMSCs replication, producing a higher number of cells in comparison to the plastic surface environment. Allogeneic or autologous blood products behave similarly. The work suggests that canine ADMSCs can be expanded in the absence of xenogeneic supplements, thus increasing the safety of cellular preparations. Furthermore, the 3D fibrin-based matrices could represent a simple, readily available environments for effective in vitro expansion of ADMSCs using allogeneic or autologous blood-products.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Xenobiotic-Free Medium Guarantees Expansion of Adipose Tissue-Derived Canine Mesenchymal Stem Cells Both in 3D Fibrin-Based Matrices and in 2D Plastic Surface Cultures

Suelzu

Conti

Khalidy

et al. 2020

Cells

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“…ePL concentrations higher than 30% showed a detrimental effect on CB-MSC proliferation rates compared to equal concentrations of FBS ( 78 ). One study assessed chondrogenic differentiation in equine BM-MSCs cultured in ePL and found no difference between ePL and FBS cultured cells ( 80 ), while another study in equine peripheral blood derived MSCs found a decrease in proliferation in cells cultured in 15% PL compared to FBS ( 40 ). In canine AD-MSCs, Hagen et al found that when cultured in PL, the cells lost their spindle-like formations and were unable to be dissociated from the culture flask ( 81 ).…”

Section: Fbs In Msc Culturementioning

confidence: 99%

A Review of Fetal Bovine Serum in the Culture of Mesenchymal Stromal Cells and Potential Alternatives for Veterinary Medicine

et al. 2022

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Fetal bovine serum (FBS) remains widely used as a supplement in cell culture media used in the isolation and expansion of mesenchymal stromal cells (MSC) despite longstanding practical, clinical, and ethical concerns over its use. As a result, research on alternative culture media supplement solutions that conserve crucial MSC characteristics has become increasingly relevant. Species-specific supplements and serum-free media such as platelet lysate or chemically defined media have been assessed for their effect in MSC cultures regarding proliferation, differentiation, and immunomodulatory capacity. While none of the alternatives offer a complete solution in replacing traditional FBS supplemented media for culturing MSCs for all species, short-term or transitional use of FBS-free media can perform equally well and could address some of the concerns over the use of FBS.

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“…Platelet lysate is generated from platelet-rich plasma (PRP) and contains high concentrations of growth factors, including transforming growth factor-β (TGF-β), platelet-derived growth factors (PDGF), and vascular endothelial growth factor (VEGF) due to degranulation of platelet alpha granules (22). Media supplementation with PL does not appear to compromise MSC proliferation, viability, or potency when compared to FBS (11,12,23,24). Importantly, several recent studies have also shown that PL has significant anti-inflammatory properties in vitro (12,20).…”

Section: Introductionmentioning

confidence: 99%

Comparing the immunomodulatory properties of equine BM-MSCs culture expanded in autologous platelet lysate, pooled platelet lysate, equine serum and fetal bovine serum supplemented culture media

et al. 2022

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Joint injury often leads to cartilage damage and posttraumatic inflammation, which drives continued extracellular matrix degradation culminating in osteoarthritis. Mesenchymal stem cells (MSCs) have been proposed as a biotherapeutic to modulate inflammation within the joint. However, concerns have been raised regarding the immunogenicity of MSCs cultured in traditional fetal bovine serum (FBS) containing media, and the potential of xenogenic antigens to activate the immune system causing rejection and destruction of the MSCs. Xenogen-free alternatives to FBS have been proposed to decrease MSC immunogenicity, including platelet lysate (PL) and equine serum. The objective of this study was to compare the immunomodulatory properties of BM-MSCs culture-expanded in media supplemented with autologous PL (APL), pooled PL (PPL), equine serum (ES) or FBS. We hypothesized that BM-MSCs culture expanded in media with xenogen-free supplements would exhibit superior immunomodulatory properties to those cultured in FBS containing media. Bone marrow-derived MSCs (BM-MSCs) were isolated from six horses and culture expanded in each media type. Blood was collected from each horse to isolate platelet lysate. The immunomodulatory function of the BM-MSCs was assessed via a T cell proliferation assay and through multiplex immunoassay quantification of cytokines, including IL-1β, IL-6, IL-8, IL-10, and TNFα, following preconditioning of BM-MSCs with IL-1β. The concentration of platelet-derived growth factor BB (PDGF-BB), IL-10, and transforming growth factor-β (TGF-β) in each media was measured via immunoassay. BM-MSCs cultured in ES resulted in significant suppression of T cell proliferation (p = 0.02). Cell culture supernatant from preconditioned BM-MSCs cultured in ES had significantly higher levels of IL-6. PDGF-BB was significantly higher in APL media compared to FBS media (p = 0.016), while IL-10 was significantly higher in PPL media than ES and FBS (p = 0.04). TGF-β was highest in APL media, with a significant difference in comparison to ES media (p = 0.03). In conclusion, expansion of equine BM-MSCs in ES may enhance their immunomodulatory abilities, while PL containing media may have some inherent therapeutic potential associated with higher concentrations of growth factors. Further studies are needed to elucidate which xenogen-free supplement optimizes BM-MSC performance.

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Autologous Platelet Lysate Does Not Enhance Chondrogenic Differentiation of Equine Bone Marrow-Derived Mesenchymal Stromal Cells Despite Increased TGF-β1 Concentration

Cited by 13 publications

References 46 publications

Xenobiotic-Free Medium Guarantees Expansion of Adipose Tissue-Derived Canine Mesenchymal Stem Cells Both in 3D Fibrin-Based Matrices and in 2D Plastic Surface Cultures

Xenobiotic-Free Medium Guarantees Expansion of Adipose Tissue-Derived Canine Mesenchymal Stem Cells Both in 3D Fibrin-Based Matrices and in 2D Plastic Surface Cultures

A Review of Fetal Bovine Serum in the Culture of Mesenchymal Stromal Cells and Potential Alternatives for Veterinary Medicine

Comparing the immunomodulatory properties of equine BM-MSCs culture expanded in autologous platelet lysate, pooled platelet lysate, equine serum and fetal bovine serum supplemented culture media

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