Research on the Regulatory Effects of Bovine Skeletal Muscle Cells on Adipocytes Under Co-Culture Conditions
[Objective]Marbling is one of the key indicators for meat quality,formed by the joint development of bovine skeletal muscle cells and bovine adipocytes.The purpose of this study was to simulate the interaction effect of bovine skeletal muscle cells and adipocytes in the process of meat formation in vivo to the greatest extent,and to establish two cell co-culture systems in vitro to explore the regulatory effects of bovine skeletal muscle cell secretion and metabolic factors on adipocytes under the co-culture system.[Method]Bovine adipocytes and skeletal muscle cells were isolated separately using tissue block and enzymatic digestion methods.Cells were identified for purity and differentiation potential by phenotypic identification and gene expression profiling,the conditioned medium exchange co-cultivation systems constructed further,and Transwell co-cultivation systems involving bovine skeletal muscle cells and bovine fat cells was conducted.The impact of secretory and metabolic products from bovine skeletal muscle cells on the proliferation and differentiation of fat cells under co-culture conditions was assessed using techniques,such as real-time quantitative PCR(qPCR),EdU staining,and Oil Red O staining.[Result]In this study,two co-culture systems of bovine skeletal muscle cells and adipocytes were successfully constructed.Under the condition of medium exchange co-culture,the expression of bovine fat cell proliferation marker genes,including PCNA,CDK2,CCNE2,and CCND1,was significantly downregulated(P＜0.01).Additionally,the expression of adipogenesis marker genes FABP4 and PPARγ was significantly decreased(P＜0.05);the expression of LPL was greatly reduced(P＜0.01).On the other hand,in the Transwell co-culture system,the expression of the bovine fat cell proliferation marker gene CCND1 was significantly downregulated(P＜0.05);the expression of PCNA,CDK1,and CCNE2 was greatly downregulated(P＜0.01).Additionally,the expression of the adipogenesis marker gene FABP4 was significantly decreased(P＜0.05),while the expression of PPARγ,C/EBPβ,and LPL was extremely significantly decreased(P＜0.01).[Conclusion]The results of this study indicated that secretory and metabolic products from bovine skeletal muscle cells could suppress the proliferation and differentiation of bovine adipocytes by inhibiting the expression of proliferation marker genes,including PCNA,CDK1,CDK2,CCNE2,and CCND1,as well as lipogenic marker genes,such as PPARγ,FABP4,CEBPβ,and LPL.The results of this study indicated that secretory and metabolic products from bovine skeletal muscle cells could suppress the proliferation and differentiation of bovine adipocytes by inhibiting the expression of proliferation marker genes,such as PCNA,CDK1,CDK2,CCNE2 and CCND1,as well as lipogenic marker genes,such as PPARγ,FABP4,CEBPβ and LPL.
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