Impact of different formulations of platelet lysate on proliferative and immune profile of equine mesenchymal stromal cells.
Abstract: Platelet lysate (PL) is investigated as a potential replacement for fetal bovine serum (FBS) in cell culture. However, there is limited research on its impact on the immune profile of equine mesenchymal stromal cells (eMSCs). This study aimed to evaluate the effects of different PL formulations on the proliferative capacity, multipotentiality, and immune profile of equine adipose tissue-derived MSCs (eAD-MSCs). growth kinetics and trilineage differentiation of eAD-MSCs ( = 7) were assessed under three culture conditions: medium-concentration PL (MPL), high-concentration PL (HPL), and FBS as a control. The immune profile was evaluated by studying the expression of immunogenic receptors such as MHC I, MHC II, and immunomodulatory molecules IL-6, IL-10, and TNF-α, determined by gene expression, surface marker expression, and cytokine quantification. Both PL formulations, pooled from 5 donors, exhibited 3.3 and 6.5-fold higher platelet counts than baseline plasma for MPL and HPL, respectively. Higher concentrations of TGF-β and PDGF were found in both PL formulations compared to baseline. Furthermore, MPL and HPL subcultures demonstrated proliferative, clonogenic, and multipotent capacities similar to FBS. The immune profile of PL-cultured cells exhibited gene expression levels related to immunogenicity and immunomodulation similar to the reference condition, and the surface antigen presence of MHC II was also similar. However, HPL media exhibited higher IL-6, IL-10, and TNF-α concentrations in the culture supernatant. In conclusion, both PL media contained higher concentrations of growth factors compared to FBS, supporting the culture of eAD-MSCs with proliferative, clonogenic, and multipotent capacity similar to the reference medium. Nonetheless, PL usage led to a variation in the immunomodulatory cytokine microenvironment, with higher concentrations of IL-6, IL-10, and TNF-α in HPL media compared to MPL and FBS.
Copyright © 2024 Yaneselli, Ávila, Rossi, Rial, Castro, Estradé, Suárez and Algorta.
Publication Date: 2024-08-05 PubMed ID: 39161460PubMed Central: PMC11330840DOI: 10.3389/fvets.2024.1410855Google Scholar: Lookup
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Summary
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The research focuses on studying the impact of different concentrations of platelet lysate (PL) on the proliferative ability and immune profile of equine adipose tissue-derived mesenchymal stromal cells (eAD-MSCs). The key finding was that both low and high concentration PL formulations promoted similar cell culture results as the commonly used fetal bovine serum (FBS), with the high concentration PL also causing increased expression of certain immunomodulatory molecules.
Introduction to the Research
- The researchers used PL as an alternative to FBS in cell culture with the aim of analysing its effects on the growth and gene expression profile of eAD-MSCs.
- The research assessed whether the level of concentration of the PL would affect proliferation, differentiation into multiple cell types, and the immune profile of cells.
Methods Employed in the Research
- The study involved the use of different PL formulations and FBS as a control. The formulations were a medium concentration of PL (MPL) and a high concentration of PL (HPL).
- The experiments analyzed the expression of immunogenic receptors (MHC I, MHC II) and immunomodulatory molecules (IL-6, IL-10, and TNF- α) using gene expression, surface marker expression, and cytokine quantification methods.
Findings of the Research
- Both PL formulations demonstrated a higher count of platelets compared to baseline plasma. Also, both MPL and HPL resulted in higher concentrations of the growth factors TGF-β and PDGF compared to baseline.
- Cells grown in both PL formulations showed a similar growth rate, the ability to generate colonies (clonogenic capacity), and the potential to differentiate into different types of cells (multipotent capacity) as those grown in FBS.
- Although the immune profile in terms of immunogenic receptors was found to be similar in both PL media and when FBS was used, the expression of immunomodulatory molecules IL-6, IL-10, and TNF-α was higher in the HPL media than the MPL and FBS cultures.
Conclusion of the Research
- PL media, irrespective of its concentration, supported the growth and multipotent capacity of eAD-MSCs similar to the culture containing FBS.
- The PL media showed higher concentrations of growth factors, particularly in HPL, which also exhibited an enhanced immunomodulatory cytokine microenvironment.
- These findings suggest that using PL as a media in cell culture of eAD-MSCs may affect the cell’s innate immune response, which necessitates further study in the context of therapeutic applications.
Cite This Article
APA
Yaneselli K, Ávila G, Rossi A, Rial A, Castro S, Estradé MJ, Suárez G, Algorta A.
(2024).
Impact of different formulations of platelet lysate on proliferative and immune profile of equine mesenchymal stromal cells.
Front Vet Sci, 11, 1410855.
https://doi.org/10.3389/fvets.2024.1410855 Publication
Researcher Affiliations
- Unidad de Inmunología e Inmunoterapia, Departamento de Patobiología, Facultad de Veterinaria, Universidad de la República, Montevideo, Uruguay.
- Unidad de Inmunología e Inmunoterapia, Departamento de Patobiología, Facultad de Veterinaria, Universidad de la República, Montevideo, Uruguay.
- Departamento de Desarrollo Biotecnológico, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay.
- Departamento de Desarrollo Biotecnológico, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay.
- Unidad de Clínica y Cirugía de Equinos, Departamento de Clínica y Hospital Veterinario, Facultad de Veterinaria, Universidad de la República, Montevideo, Uruguay.
- Unidad de Reproducción Animal, Departamento de Producción Animal y Salud de los Sistemas Productivos, Facultad de Veterinaria, Universidad de la República, Montevideo, Uruguay.
- Unidad de Farmacología y Terapéutica, Departamento de Clínicas y Hospital Veterinario, Facultad de Veterinaria, Universidad de la República, Montevideo, Uruguay.
- Unidad de Inmunología e Inmunoterapia, Departamento de Patobiología, Facultad de Veterinaria, Universidad de la República, Montevideo, Uruguay.
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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