Study On Inducing Differentiation Of Bone Marrow Mesenchymal Stem Cells Into Thermogenic Adipocytes

Objective: To study the potential of C57BL6/J mice and adults BMSCs to differentiate into thermogenic adipocytes,and provide more possibilities for the source of thermogenic adipocytes in the body.Methods: 1.To compare the ability of inducing mice BMSCs to differentiate into thermogenic adipocytes under different conditions and intervention factors by detecting the amount of gene expression: a.mice BMSCs of different ages: 6 weeks,10-12 weeks,16-20 weeks old;b.the influence of rosiglitazone during the induction of adipogenesis;c.proximal and distal long bone BMSCs;d.the intervention of c AMP agonists on BMSC-derived adipocytes.2.Mice and adults BMSCs adipogenic induction time gradient experiment: detect the expression of thermogenic genes at different time points of the induction process.3.Prdm16-overexpressed lentivirus were packaged and transfected into mice BMSCs,which were induced adipogenesis to detect the expression of related genes such as thermogenesis.4.Cold stimulation experiment: two groups of mice were placed in 15?and 23?environment for one week.BAT and inguinal WAT were extracted for HE staining and BMSCs were extracted to detect the expression of thermogenic genes after adipogenesis.5.After c AMP agonists interfered with adipocytes derived from adults BMSCs,the expression of thermogenesis,adipogenesis and lipolysis-related genes were detected;the TG content in the cells and in the culture medium were detected;the changes in the morphology and number of adipocytes under a microscope were observed.6.The samples of adults BMSCs before,after adipogenic induction and then treated with c AMP agonist were sent to the company for transcriptome sequencing and analysis of the results.Results: 1.The effect of adipogenic differentiation of BMSCs in 16-20 weeks old mice was better,and the intervention of rosiglitazone helped to increase the expression of the key gene Pgc1? for thermogenesis.2.Proximal BMSCs had better adipogenic differentiation,higher expression of Pgc1? and more potential to differentiate into thermogenic adipocytes.3.With the prolonged adipogenic induction time of mice and adults BMSCs,the expression of thermogenic genes gradually increased.4.The c AMP agonist promoted the expression of thermogenic genes of adipocytes derived from mice and adults BMSCs,and initiated the lipolysis process.5.Prdm16 expression did not change significantly during the adipogenesis induction process of mice and adults BMSCs;After overexpressing Prdm16 in mice BMSCs and inducing adipogenesis,the expression of thermogenic genes did not change significantly.6.RNA-seq results showed that the induction of adults BMSCs adipogenesis and the intervention with c AMP agonist both activated thermogenesis and cell migration related pathways.Conclusion: Both mice and adults BMSCs can differentiate into thermogenic adipocytes;Proximal BMSCs in mice were more capable of differentiating into thermogenic adipocytes.