Cell and Tissue Journal

Abstract Aim: An endogenous repairman of the central nervous system is possible via neural stem cells. Some researchers have suggested the neural protective of some medicinal herbs such as; Urtica dioica. The purpose of this study is creating an oxidative condition in the neural stem cell cultures and then treated the cells with methanolic extract of nettle leaves in vitro.
Material and Methods: In this experimental study, the hippocampal neural stem cells extraction was performed by enzymatic digestion in newborn rats. The verification of these cells as nerve cells was carried out by the morphological and immunocytochemistry examinations. Before treatment, the cells were exposed for 24 hours to oxidative stress condition and then nettle leaf extract was added to the cell plates at of 2.5, 5, 10, 20 and 40 μg/ml and cells were maintained for 24 hours, separately. The final evaluation of the cell proliferation was performed by MTT viability test. The groups included; an experimental group that was treated with extract and the control group.
Results: Neural stem cells had neural morphology and expressed nestin marker. Proliferation percentage of the neural stem cells was higher in the treatment groups than the control group. In addition, MTT assay results showed that the percentage of live cells in the treated groups increased, as the proliferation of the neural stem cells significantly increased (p < 0/05) at 20μg/ml.
Conclusion: Methanolic extract of nettle leaf have neuroprotective effects and could adjust oxidative stress condition in vitro. It was able to improve the dysfunction of the central nervous system, following the production of free radicals.
 

Abstract Aim: In current study capability of differentiation of embryonic carcinoma cells P19 to cardiomyocyte through inducing effects of DMSO and 5-Azacytidin individually or in combinative form in laboratory condition were examined. Material and Methods: In order to differentiate induction of P19, Embryoid Bodies (EBs) formed through hanging drops method during two days. Then EBs have induced by 2 μM of 5-Azacytidin (5-Aza group), 0/5% DMSO  (DMSO group) or both 5-Azacytidin and DMSO (Aza+DM Group) during ten more days. During differentiation beating number per minute counted every two days in all groups by invert Microscopy. Gene expression such as:  α-Myosin Heavy Chain (α-MHC) and Myosin Light Chain (MLC) and Connexin-43 have examined by RT-PCR in last day. To more accuracy, F-actin protein expression also illustrated through immunocytochemistry method. Results: Morphological changes in differentiated cardiomyocytes derived from P19 illustrated in all groups. However the most changes such as significant increasing in size and number of process, their branches and attachment between branches happened in Aza+DM group. Daily beating counter per minute revealed increasing in beating number from 2+2 to 2+8 in all groups. RT-PCR analysis also revealed expression of both α-MHC and MLC in Aza+DM Group. However, F-action has expressed only in Aza+DM group. Conclusion: The Results have shown combination of two 5-Azacytidin and DMSO inducer caused effective cell culture differentiate of P19 to cardiomyocyte.

Abstract Aim:Recently, attention has been diverted to the pluripotency characteristic of Spermatogonia Stem Cells (SSCs) in in-vitro culture. The aim of this study was to evaluate the morphological and molecular genetics changes of differentiated SSCs to Embryonic-like Stem cell (ESC). Material and Methods: The SSCs were collected from neonatal mouse testis via a two-step mechanical and enzymatic digestion and cultured in DMED containing 15% FBS.  ES-like cells colonies was appeared in the fifth passage. The expression of pluripotency and spermatogonia specific genes; Nanog, SOX2, Klf4, Stra8, DAZL, Plzf as well as the expression of puripotency markers Oct4 and Alkaline Phosphatase Activity (ALP) of ESC respectively was investigated using RT-PCR and immunocytochemical techniques and compared with ESCs. Results: The pluripotent characteristic of ES-like cells derived from SSCs was confirmed based on morphological investigation and high expression of ALP as well as pluripotency marker Oct4. The expression of specific spermatogonia genes like Stra8, DAZL, Plzf was decreased and the expression of pluripotency genes such as  Nanog, SOX2, Klf4  was increased during the differentiation of SSCs to ES-like cells. Conclusion: In vitro culture of SSCs was conformed to induced ES-like cells. This process was accompanied by wide alteration in molecular profile expression of specific spermatogonia and pluripotency genes. In over all, SSCs can be considered as an opening window to generate pluripotent stem cell.