Author/Authors :
Abouhamzeh، Beheshteh نويسنده Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran , , Salehi، Mohammad نويسنده Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran , , Hosseini، Ahmad نويسنده Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran , , Masteri-Farahani، Ali Reza نويسنده Department of Cell Biology and Anatomical Sciences, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran , , Fadai، Fatemeh نويسنده Department of Cell Biology and Anatomical Sciences, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran , , Heidari، Mohammad Hasan نويسنده Department of Biology and Anatomical Sciences, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran. , , Nourozian، Mohsen نويسنده Department of Biology and Anatomical Sciences, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran , , Soleimani، Masoud نويسنده , , Khorashadizadeh، Mohsen نويسنده Department of Medical Biotechnology, School of Advanced Medical Technology, Tehran University of Medical Sciences, Tehran, Iran , , Mossahebi-Mohammadi، Majid نويسنده Department of Stem Cell Biology, Stem Cell Technology Research Center, Tehran, Iran , , Mansouri، Ardalan نويسنده Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran ,
Abstract :
Objective: Many studies have focused on the epigenetic characteristics of donor cells
to improve somatic cell nuclear transfer (SCNT). We hypothesized that the epigenetic
status and chromatin structure of undifferentiated bovine adipose tissue-derived stem
cells (BADSCs) would not remain constant during different passages. The objective of
this 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-4
expression (OCT4) and acetylation of H3K9 (H3K9ac) in BADSCs cultures and different
passages in vitro.
Materials and Methods: In this experimental study, subcutaneous fat was obtained
from adult cows immediately post-mortem. Relative level of DNMTs and HDACs was
examined using quantitative real time polymerase chain reaction (q-PCR), and the
level 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 decrease
in its level was seen at P7. The highest and lowest levels of H3K9ac were observed
at P5 and P7, respectively. At P5, the expression of HDACs and DNMTs was significantly
decreased. In contrast, a remarkable increase in the expression of DNMTs was
observed at P7.
Conclusion: Our data demonstrated that the epigenetic status of BADSCs was variable
during culture. The P5 cells showed the highest level of stemness and multipotency and
the lowest level of chromatin compaction. Therefore, we suggest that P5 cells may be
more efficient for SCNT compared with other passages.
