Effects of Muse stem cells on gene expression differences of nucleus pulposus cells induced by lipopolysaccharide
Objective To investigate the impact of Muse(Multilineage differentiating stress endur-ing cells)stem cells on transcriptome gene expression variations in lipopolysaccharide(LPS)-induced nu-cleus pulposus cells(NPCs).Methods Muse stem cells,derived from human umbilical cord mesenchy-mal stem cells through trypsin digestion,were co-cultured with LPS-stimulated rat primary NPCs to simulate intervertebral disc degeneration.Muse stem cells and mesenchymal stem cells were co-cultured with LPS-stimulated NPCs for 24 h.Muse stem cells were verified through cell immunofluorescence using SSEA-3 and CD105 markers,while rat primary NPCs were identified by assessing Collagen Ⅱ and Aggrecan expres-sion.A total of 3 samples each of untreated NPCs,LPS-stimulated NPCs,and NPCs co-cultured with Muse stem cells and MSCs were subjected to total RNA extraction.Transcriptome sequencing was then conduc-ted,and the obtained gene expression data were differentially analyzed using the DESeq2 package in R soft-ware.Bioinformatics methods were employed to identify differentially expressed genes(DEGs).Subse-quently,gene ontology(GO)analysis and kyoto encyclopedia of genes and genomes(KEGG)analysis were performed on these genes to identify the potential roles of Muse stem cells in the progression of interverte-bral disc degeneration(IDD).Results Following LPS stimulation,NPCs exhibited significant gene ex-pression changes compared to untreated cells,with 1 560 up-regulated DEGs and 2 173 down-regulated DEGs.Co-culture with Muse stem cells resulted in 143 up-regulated DEGs and 62 down-regulated DEGs.Similarly,co-culture with mesenchymal stem cells(MSCs)led to 142 up-regulated DEGs and 72 down-reg-ulated DEGs.GO database analysis highlighted enrichment in positive regulation of ERK1 and ERK2 cas-cades,extracellular matrix(ECM)formation,and cell migration.KEGG pathway analysis identified signif-icant pathways such as ECM receptor interaction,TGF-β signaling pathway,cytokine-receptor interaction,tumor necrosis factor(TNF)signaling pathway,glycosaminoglycan biosynthesis,phosphatidylinositol 3 ki-nase(PI3K)-protein kinase B(Akt)signaling pathway,chemokine signaling pathway,and calcium signa-ling pathway.Conclusion ECM components,growth factors,collagen fibrin constituents,inflammatory chemokines,and TNF and PI3K-Akt signaling pathways play pivotal roles in Muse stem cells'repair of damaged NPCs.
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