Functonal analyss exposed that only a smaller percentage of TRAF MSCs were postve for O Red O stanng versus AL cells.Addtonally, only few TRAF MSCs had been in a position to keep glycogeand uptake low densty lpoproten, com pared wthhL cells.Notably, TRAF MSCs have been stl in a position to even more dfferent ate, below 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 carried out a comparatve analyss for that plurpotency markers Oct four, Sox 2 and Nanog TRAF MSCs, AF MSCs, AL andhL cells by Genuine tme PCR analyss.AF MSCs and TRAF MSCs exhbtedhgher levels of expressofor Oct 4, Sox 2 and Nanog compared wth AL andhL cells, documentng the prmtve phenotype within the former cells.
addton, the lysosomal actvty and mtochodral mass of TRAF MSCs determned by LysoTracker and MtoTracker uorescent dyes, respectvely, STA-9090 888216-25-9 was smar to AF MSCs.however, AL cells andhL cells exhbted sgncantly reduced ratos of mtochondra lysosomal uores cence 0.520.04 and 0.310.04, respectvely.Moreover, semquanttatve PCR analyss unveiled that TRAF MSCs dd not express any of the adpogenc markers, like fatty acd bndng prote4, PPARg and LPL.Thehepatogenc marker tyrosne amnotransferase was expressed smar levels AF MSCs and TRAF MSCs, whch mples that later cell typeshave returned to a additional 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 three cell forms expressed Oct 4, Sox 2 and Nanog at protelevel, as demonstrated by mmunouorescent stanng.Moreover, we carried out a proteomc analyss in the AF, DAF and TRAF MSCs, dentfyng dfferentally expressed protens by 2DE gel and MS.A comparsoof the expressolevels from the respectve selleck chemical protens was establshed usng PDQuest 8 application.Protens dfferentally expressed 41.5 fold the three populatons, are descrbed deta Supplementary Table 2.Spec cally, the metabolc proteXaa Professional dpeptdase 28 was identified for being expressed hgh degree n AF MSCs compared wth TRAF MSCs.Also, Rho GDdssocaton nhbtor 1 was expressed hgher amounts DAF MSCs and TRAF MSCs compared wth AF MSCs.GDR1 s reported to partcpate the approach of dedfferentatoof smooth muscle cells.29 addton, TRAF MSCs expressed hgh levels the elongatofactor 1 delta and perox redox4, both connected to dfferentaton of MSCs.
30,31 To dene the dfferences the prole in the molecular medators secreted by AF, DAF and TRAF MSCs, we examned the condtoned medum derved from AF MSCs and drectly in contrast ths wth DAF MSC CM and TRAF MSC CM, respectvely, usng proteome prolerhumaarrays.The array analyss allowed the dentcatoof twenty secreted development components for AF MSCs, 16 for DAF MSCs and 25 for TRAF

MSCs.