AUTHOR=Zhai Zhu-Hui , Li Jun , You Zhao , Cai Yang , Yang Jie , An Jie , Zhao Di-Peng , Wang He-Jie , Dou Min-Min , Du Rong , Qin Jian 

TITLE=Feline umbilical cord-derived mesenchymal stem cells: isolation, identification, and antioxidative stress role through NF-κB signaling pathway

JOURNAL=Frontiers in Veterinary Science

VOLUME=10

YEAR=2023

URL=https://www.frontiersin.org/journals/veterinary-science/articles/10.3389/fvets.2023.1203012

DOI=10.3389/fvets.2023.1203012

ISSN=2297-1769

ABSTRACT=<p>At present, the differentiation potential and antioxidant activity of feline umbilical cord-derived mesenchymal stem cells (UC-MSCs) have not been clearly studied. In this study, feline UC-MSCs were isolated by tissue adhesion method, identified by flow cytometry detection of cell surface markers (CD44, CD90, CD34, and CD45), and induced differentiation toward osteogenesis and adipogenesis <italic>in vitro</italic>. Furthermore, the oxidative stress model was established with hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>) (100 μM, 300 μM, 500 μM, 700 μM, and 900 μM). The antioxidant properties of feline UC-MSCs and feline fibroblasts were compared by morphological observation, ROS detection, cell viability via CCK-8 assay, as well as oxidative and antioxidative parameters via ELISA. The mRNA expression of genes related to NF-κB pathway was detected via quantitative real-time polymerase chain reaction, while the levels of NF-κB signaling cascade-related proteins were determined via Western Blot. The results showed that feline UC-MSCs highly expressed CD44 and CD90, while negative for CD34 and CD45 expression. Feline UC-MSCs cultured under osteogenic and adipogenic conditions showed good differentiation capacity. After being exposed to different concentrations of H<sub>2</sub>O<sub>2</sub> for eight hours, feline UC-MSCs exhibited the significantly higher survival rate than feline fibroblasts. A certain concentration of H<sub>2</sub>O<sub>2</sub> could up-regulate the activities of SOD2 and GSH-Px in feline UC-MSCs. The expression levels of <italic>p50</italic>, <italic>MnSOD</italic>, and <italic>FHC</italic> mRNA in feline UC-MSCs stimulated by 300 μM and 500 μM H<sub>2</sub>O<sub>2</sub> significantly increased compared with the control group. Furthermore, it was observed that 500 μM H<sub>2</sub>O<sub>2</sub> significantly enhanced the protein levels of p-IκB, IκB, p-p50, p50, MnSOD, and FHC, which could be reversed by BAY 11-7,082, a NF-κB signaling pathway inhibitor. In conclusion, it was confirmed that feline UC-MSCs, with good osteogenesis and adipogenesis abilities, had better antioxidant property which might be related to NF-κB signaling pathway. This study lays a foundation for the further application of feline UC-MSCs in treating the various inflammatory and oxidative injury diseases of pets.</p>