Functonal analyss unveiled that only a minor percentage of TRAF MSCs had been postve for O Red O stanng versus AL cells.Addtonally, only number of TRAF MSCs were able to store glycogeand uptake lower densty lpoproten, com pared wthhL cells.Notably, TRAF MSCs have been stl capable to even more dfferent ate, underneath approprate culture condtons nto AL and OL cells.TRAF MSCs exhbted smar characterstcs to AF MSCs.To obtafurther nsght nto the modifications the gene expressoprole durng transdfferentaton, we performed a comparatve analyss to the plurpotency markers Oct 4, Sox 2 and Nanog TRAF MSCs, AF MSCs, AL andhL cells by Actual tme PCR analyss.AF MSCs and TRAF MSCs exhbtedhgher amounts of expressofor Oct four, Sox two and Nanog in contrast wth AL andhL cells, documentng the prmtve phenotype from the former cells.
addton, the lysosomal actvty and mtochodral mass of TRAF MSCs determned by LysoTracker and MtoTracker uorescent dyes, respectvely, kinase inhibitor library for screening was smar to AF MSCs.even so, AL cells andhL cells exhbted sgncantly reduce ratos of mtochondra lysosomal uores cence 0.520.04 and 0.310.04, respectvely.Also, semquanttatve PCR analyss uncovered that TRAF MSCs dd not express any with the adpogenc markers, such as fatty acd bndng prote4, PPARg and LPL.Thehepatogenc marker tyrosne amnotransferase was expressed smar ranges AF MSCs and TRAF MSCs, whch mples that later on cell typeshave returned to a even more prmtve stem cell lke stage.AF MSCs, DAF MSCs and TRAF MSCs exhbt smar clonogenc potental and proteome prole.AF, DAF and TRAF MSCs shared smartes clonogenc potental, as determned by colony formng unt broblast assay.
addton, all 3 cell sorts expressed Oct four, Sox 2 and Nanog at protelevel, as demonstrated by mmunouorescent stanng.Furthermore, we carried out a proteomc analyss of the AF, DAF and TRAF MSCs, dentfyng dfferentally expressed protens by 2DE gel and MS.A comparsoof the expressolevels from the respectve supplier FK866 protens was establshed usng PDQuest eight software program.Protens dfferentally expressed 41.five fold the three populatons, are descrbed deta Supplementary Table two.Spec cally, the metabolc proteXaa Pro dpeptdase 28 was observed to get expressed hgh degree n AF MSCs compared wth TRAF MSCs.Additionally, Rho GDdssocaton nhbtor 1 was expressed hgher amounts DAF MSCs and TRAF MSCs compared wth AF MSCs.GDR1 s reported to partcpate the procedure of dedfferentatoof smooth muscle cells.29 addton, TRAF MSCs expressed hgh levels the elongatofactor 1 delta and perox redox4, each linked to dfferentaton of MSCs.
30,31 To dene the dfferences the prole of the molecular medators secreted by AF, DAF and TRAF MSCs, we examned the condtoned medum derved from AF MSCs and drectly compared ths wth DAF MSC CM and TRAF MSC CM, respectvely, usng proteome prolerhumaarrays.The array analyss allowed the dentcatoof twenty secreted development elements for AF MSCs, 16 for DAF MSCs and 25 for TRAF

MSCs.