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[Display omitted] • Effects of three flavonoids on porcine muscle stem cells were investigated. • 3,2′-DHF and QUE had better pro-proliferation and pro-differentiation, respectively. • The optimized combination of flavonoids were used in the process of cultured meat. • The contraction area of cultured meat were promoted by QUE. Stemness decline of muscle stem cells (MuSCs) is a significant problem in cultured meat processing. In the present study, three flavonoids (quercetin, icariin, and 3,2′-dihydroxyflavone) with multi concentrations were evaluated to promote the proliferation and differentiation of porcine muscle stem cells. In the proliferation phase, 3,2′-dihydroxyflavone (10 μM) significantly amplified the cells by 34% and up-regulated the expression of paired box transcription factor 7 (PAX7) by 60%, which was higher than quercetin (75 nM) and icariin (7.5 nM). In the differentiation phase, quercetin (50 nM) showed the best pro-differentiation effect and up-regulated the expression of myosin heavy chain (MYHC) by 4.73-fold compared with the control group. These results indicated that flavonoids had a significant impact on promoting the proliferation and differentiation of porcine MuSCs, and 3,2′-dihydroxyflavone (10 μM) for proliferation and quercetin (50 nM) for differentiation were the optimal combinations. [ABSTRACT FROM AUTHOR]