Figure 6. Experimental design

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Equine adipose tissue-derived mesenchymal stem cells (EqAT-MSCs) (P3) were primed with 5 µM of cannabidiol (CBD)-rich cannabis extract + complete culture medium and the other group was primed with 7 µM of CBD-rich cannabis extract + complete culture medium. EqAT-MSCs cultured in a complete culture medium were used as the control. After 24 h of priming, the morphological evaluation and gene expression analysis of cytokines such as interleukin 1 beta (IL-1β), interleukin 6 (IL-6), interleukin 10 (IL-10), interferon-gamma (IFN-γ), and tumor necrosis factor-alpha (TNF-α) were performed. Cell metabolic activity was assessed using the MTT assay, and cellular senescence was evaluated through β-galactosidase activity. The apoptosis assay was carried out using annexin and 7-AAD. In addition to the evaluation of cannabinoid receptors 1 (CB1) and 2 (CB2) gene expression in naïve EqAT-MSCs (P2–P5).

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Created in BioRender. Amorim, R. (2025) https://app.biorender.com/citation/6745ff5939d174eac4087843
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Amorim, R. (2025). Figure 6. Experimental design. Created in BioRender. https://app.biorender.com/citation/6745ff5939d174eac4087843

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