Evaluation of Major Histocompatibility Complex Genes Expression in Somatic Cell Nuclear Transfer Embry-onic Stem Cells

Background: Embryonic stem cells (ESCs) have the potential to differentiate into almost all cell types providing a promis-ing cell source in regenerative medicine. The low efficiency of somatic cell nuclear transfer (SCNT), the technique to achieve blastocyst for ESC generation, has been improved by trichos-tatin A (TSA) a histone deacetylase (HDACi) inhibitor, which change pluripotent mediated genes expression during repro-gramming. Since TSA changes MHC molecules expression on tumor cells which cause immune responses against them. Hence, this study aims to determine whether TSA affects MHC expression levels of ESCs.Materials and Methods: Toward this goal, we generated SCNT-ESCs in the presence and absence of TSA. IVF-ESCs was established as the control group. ESCs were confirmed by alkaline phosphatase assay and immunocytochemistry (ICC). The expression level of MHC molecules (i.e. H2Kb, H2Kd,H2Dd, Qa-1) for both blastocyst and ESCs generated by two techniques were examined by qRT-PCR.Results: Our results revealed that the expression levels of H2Kb, H2Kd, H2Dd, and Qa-1 (MHCI) in SCNT-ESCs in-creased in the presence of TSA compared to control group but they were not statistically significant. In contrast, these genes were significantly downregulated in the TSA negative group. We observed increased expression level of MHCI molecules in both blastocyst-derived TSA positive and negative SCNT.Conclusion: Although TSA plays a key role in the develop-mental rate of embryos as well as establishment of ESC lines after SCNT, behind of its negative effect on MHC expression we suggest eliminating TSA treatment after SCNT, so that we could gain better NT-ESC.

کلیدواژه ها:

Somatic Cell Nuclear Transfer ، Major Histocompat-ibility Complex ، Stem Cell