DNA Methylation and Histone Acetylation Patterns in Cultured Bovine Adipose Tissue-Derived Stem Cells (BADSCs)

Objective: many studies have focused on the epigenetic characteristics of donor cellsto improve somatic cell nuclear transfer (scnt). we hypothesized that the epigeneticstatus and chromatin structure of undifferentiated bovine adipose tissue-derived stemcells (badscs) would not remain constant during different passages. the objective ofthis study was to determine the mrna expression patterns of dna methyltransferases(dnmt1, dnmt3a, dnmt3b) and histone deacetyltransferses (hdac1, hdac2, hdac3)in badscs. in addition, we compared the measured levels of octamer binding protein-4expression (oct4) and acetylation of h3k9 (h3k9ac) in badscs cultures and differentpassages in vitro.materials and methods: in this experimental study, subcutaneous fat was obtainedfrom adult cows immediately post-mortem. relative level of dnmts and hdacs wasexamined using quantitative real time polymerase chain reaction (q-pcr), and thelevel of oct4 and h3k9ac was analyzed by flow cytometry at passages 3 (p3), 5(p5) and 7 (p7).results: the oct4 protein level was similar at p3 and p5 but a significant decreasein its level was seen at p7. the highest and lowest levels of h3k9ac were observedat p5 and p7, respectively. at p5, the expression of hdacs and dnmts was significantlydecreased. in contrast, a remarkable increase in the expression of dnmts wasobserved at p7.conclusion: our data demonstrated that the epigenetic status of badscs was variableduring culture. the p5 cells showed the highest level of stemness and multipotency andthe lowest level of chromatin compaction. therefore, we suggest that p5 cells may bemore efficient for scnt compared with other passages.