Gene Expression Analysis of SOX۲, NANOG, KLF۴, OCT۴, and REX۱ Genes in Cord Blood Mononuclear Cells Treated with External Stresses

Background: Induced pluripotent stem cells (iPSCS) can be obtained from autologous cells for therapeutic purposes. So, far, many studies have been done to produce induced pluripotent cells by transferring specific pluripotency genes using different methods. In this study, pluripotency gene expression induced by external stresses was assessed in cord blood mononuclear cells.Methods: In this experimental study, mononuclear cells were isolated from umbilical cord blood. Isolated cells were divided into three groups. The first group had been exposed to HCL (pH ۵.۷) for ۲۵ minutes and then transferred to the medium with normal pH. The second group was triturated with hamilton syringe for ۱۵ min (external pressure), and the last group was considered the control group and did not receive treatment. Then, total RNA was extracted on Day ۷. Gene expression of OCT۴, SOX۲, NANOG, REX۱, and KLF۴ was evaluated using qRT-PCR.Results: Gene expression of OCT۴, NANOG, REX۱, and KLF۴ was increased after exposure to acidic pH and external pressures in comparison with control cells (P < ۰.۰۵). SOX۲ gene expression was decreased in cells exposed to acidic pH but increased by external pressure.Conclusions: Exposure of umbilical cord blood mononuclear cells to acidic pH and external pressure lead to re-activation of pluripotency genes in mature cells. These findings indicate that mature cells may be reprogrammed with manipulation of environmental conditions.Background: Induced pluripotent stem cells (iPSCS) can be obtained from autologous cells for therapeutic purposes. So, far, many studies have been done to produce induced pluripotent cells by transferring specific pluripotency genes using different methods. In this study, pluripotency gene expression induced by external stresses was assessed in cord blood mononuclear cells. Methods: In this experimental study, mononuclear cells were isolated from umbilical cord blood. Isolated cells were divided into three groups. The first group had been exposed to HCL (pH ۵.۷) for ۲۵ minutes and then transferred to the medium with normal pH. The second group was triturated with hamilton syringe for ۱۵ min (external pressure), and the last group was considered the control group and did not receive treatment. Then, total RNA was extracted on Day ۷. Gene expression of OCT۴, SOX۲, NANOG, REX۱, and KLF۴ was evaluated using qRT-PCR. Results: Gene expression of OCT۴, NANOG, REX۱, and KLF۴ was increased after exposure to acidic pH and external pressures in comparison with control cells (P < ۰.۰۵). SOX۲ gene expression was decreased in cells exposed to acidic pH but increased by external pressure. Conclusions: Exposure of umbilical cord blood mononuclear cells to acidic pH and external pressure lead to re-activation of pluripotency genes in mature cells. These findings indicate that mature cells may be reprogrammed with manipulation of environmental conditions.

کلیدواژه ها:

Reprogramming ، External pressure ، Acidic pH ، Mononuclear cells ، Cord blood.