Objective: Embryonic stem CELLs (ESC) are defined by two main properties of self-renewal and their multipotency to differentiate into virtually all CELL types of the body, including endothelial CELLs. ESCs have been widely regarded as an unlimited source of CELLs in regeneration medicine and also an ideal in vitro model to investigate complex developmental processes. Here, we report a simple and efficient in vitro model to derive a nearly pure population of endothelial CELLs from a murine ESC LINE.Material and Methods: CCE ES CELLs are exposed to alpha-MEM medium containing 10% FBS for 4 days and then cultured in endothelial basal-2 medium containing vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), insulin-like growth factor (IGF), epidermal growth factor (EGF), and 2% FBS for 42 days.Results: The CELLs acquired a relatively uniform endothelial CELL morphology and were able to propagate and expand in culture. When murine ES CELL–derived endothelial CELLs (MESDECs) were cultured on Matrigel and incubated for 48 h, vessel-like tube structures consisting of CD31 (PECAM-1) or BS-1 immunoreactive CELLs were developed. Immunocytochemistry and RTPCR analyses revealed that MESDECs express endothelial CELL-specific marker proteins such as Flk-1, PECAM-1, Tie- 1, and Tie-2, in which the expressions persist for long periods of time after differentiation. The CELLs were also capable of taking up acetylated low-density lipoprotein (LDL) in culture.Conclusion: Our data suggest that MESDECs could provide a suitable in vitro model to study molecular events involved in vascular development and open up a new therapeutic strategy in regeneration medicine of cardiovascular disorders

Introduction: Embryoid bodies (EBs) are primitive embryonic structures derived from differentiating embryonic stem CELLs (ESCs). Many techniques have been used to obtain EBs, the improvement of the technique of EBs formation can help in achieving better results in ESCs differentiation into neurons, myocardiocytes, hemopoietic CELLs and others. In this investigation Sigmacote was used as a hydrophobic substrate to improve EBs formation.Material and Methods: Petri dishes` floor was been siliconized by a three-step procedure and with sigmacote. Then CCE and p19 CELL LINE were used to obtain EBs in these siliconized petri dishes. These CELLs were transmitted to petri dishes, the concentration of them was 1´105 CELLs/ml, after 48 hours embryoid bodies were counted. Control group was bacteriologic petri dishes with same conditions. Results: The results of EBs calculation showed that siliconized petri dishes in comparison with bacteriological petri dishes can be produced EBs with high efficiency and statistically it was significant (p<0.05).Conclusion: Results of this research indicated that sigmacote as a hydrophobic substrate can improve EBs formation by an easy and reproducible method. Therefore it is suggested to increase EBs formation and differentiation of ESCs.

Objective: Prostate cancer is the second cause of cancer-associated death in men. In recent years, targeted therapy for cancer has attracted the attention of researchers.Targeted therapy leads to a decrease in drug adverse effects. Studies indicate that targeting peptides for cancer CELLs represent valuable tools for diagnostics and therapeutics. Recently, phage display peptide libraries have been used to identify target peptides to a variety of cancer CELLs. In the current study, we aim to isolate peptides that target PC3 CELLs (human prostate adenocarcinoma CELLs).Methods: Four rounds of subtractive panning on control CELLs that included 5637 (bladder), Huh-7 (liver), SW480 (colon), AGS (stomach) and human fibroblast normal in addition to four rounds of positive panning on PC3 (target CELL) were performed.Polyclonal phage ELISA was used to evaluate the process of enrichment during biopanning. Subsequently, phage clones were randomly selected from titer plates, amplified by plaque-PCR, and their genomic DNA was sequenced. We conducted bioinformatic analysis for further characterization of the isolated peptides.Results: Several rounds of panning resulted in the enrichment of a number of peptides.The results of polyclonal phage ELISA indicated that the biopanning process was suCCEssful. In silico analysis showed the presence of several consensus amino acid motifs in the peptides.Conclusion: The peptides identified through biopanning can be considered as potential specific binders to PC3 CELLs. Peptides with specificity binding to target CELLs can be used for targeted gene and drug delivery to malignant tumor CELLs. Further analyses of these peptides are required to show their capacity for targeted delivery of various genes and drugs into prostate cancer CELLs.

Introduction & Objective: Previous studies have shown that ES CELLs could be induced to differentiate into neurons and gelia in vitro. Induction protocols are based on culture in the presence of an inducer such as RA. In this study, the effect of deprenyl on the differentiation of embryonic stem CELLs (ES) CELLs to neuron-like CELLs was investigated. Deprenyl is a type B monoamine oxidase inhibitor which has been used for treatment of Parkinson's disease.The drug protects neurons in a variety of modes, by inducing neurotrophin expression, it causes neuritis outgrowth. These raised the possibility of a direct effect of deprenyl on pleuripotent ES to develop into neurons.Materials & Methods: CELLs of an embryonic stem CELL LINE (CCE) were cultured as aggregates (embryoid bodies) then, they were exposed to a concentration of 10-8M deprenyl. CCE was used to evaluate the induction of neuronal differentiation by deprenyl. Morphological and immunohistochemical techniques were used to evaluate the differentiation of CCE CELLs In addition to the Cresyl violet used for morphological study, antinestin antibody was applied to characterize the neuroepithelial differentiation. Moreover, anti-synaptophysin and anti-tyrosine hydroxylase antibodies were used to characterize the neuronal phenotype of ESCs.Results: The results indicated that the concentration of 10-8M deprenyl could induce the differentiation of ESCs into neurons.

Purpose: Parkinson's disease is a degenerative disease produced by the death of dopaminergic neurons, and the response to current treatments is varied. It is important to develop a model for the evaluation of ES CELLs as an alternative model for treatment.Materials and Methods: The model for PD was developed in rats. First, ES CELLs were transplanted into experimental models in three groups: treated by RA, non-treated by RA and transfected by BDNF gene; the fourth group received culture media. Then the ability of the CELLs to improve Parkinson's disease was evaluated.The CELLs were labeled by Brdu. Histological and immunohistochemical evaluations were done at day 5, and behavioral evaluation was carried out at week 8th after transplantation.Results: At day 15, histological evaluation by H&E and crystal violet stainings indicated that the transplanted CELLs had differentiated into neural CELLs and integrated into the host tissue. Examination of transplanted CELLs with the electron microscope showed, neurons and oligodendrocyte. In addition, the Immunohistochemical result was positive for tyrosine hydroxylase, which is an important marker for. dopaminergic neurons, and confirmed the differentiation of transplanted CELLs to dopaminergic neurons. At day 5, the embryonic stem CELLs that were labeled with Brdu and expressed SSEA1 antigen were detected in the transplantation area. At week 8th after transplantation, the results of histological and Immunohistochemical investigation were confirmed by behavioral test, indicating improvement in three groups, i.e., treated by RA, non-treated by RA and transected by BDNF gene when compared with the control group.Conclusion: Our results indicated that CCE embryonic stem CELLs are an appropriate CELL LINE for transfection and differentiation to dopaminergic neurons, which can be used for research based on CELL and gene therapy.

Background and purpose: During recent years, nitric oxide (NO) has been considered as a molecule involved in migraine headaches. This free radical involves in initiation of migraine headaches via NO/cGMP signaling pathway and vascular relaxation specially big intracranial arteries. Therefore, we studied the effects of aqueous extract of Origanum vulgare and Melilotus officinalis prescribed in migraine treatment in traditional & modem medicine, on NO level in cultured endothelial CELLs.Materials and Methods: Each crude herb (25g) was mixed with 200 ml distilled water. End extract obtained after filtering and drying. Endothelial CELLs propagated in DMEM medium containing 10% FCS and 1-2% penicillin-streptomycin. The nitrite concentration was measured as an indicator of nitric oxide production according to the Griess reaction and with ELISA in 540nm.Results: Concentrations of 100, 200 and 400 µg/ml of Origanum vulgare, reduced NO levels compared with control to 13.1 % (p<0.05), 25.8% (p<0.01) and 33.9% (p<0.001) respectively. However, despite our expectation melilotus officinalis increased NO level. The concentrations of 100, 200 and 400 µg/ml of the herb, increased NO levels to 12.7 (p<0.05), 36.5% (p<0.001) & 72.9% (p<0.001) respectively.Conclusion: We concluded that aqueous extract of Origanum vulgare probably decreases migraine headaches by reducing NO and aqueous extract of Melilotus officinalis does not act through this mechanism.

Introduction: Unlimited self renewal potential of embryonic stem CELLs in differentiation to different types of CELLs Provides CELLs which can be an attractive doner source for developmental studies, CELL therapy and gene therapy sterategies. In this  research  ES CELLs  were differentiated into neuron-like CELLs. Material and Methods: At first, dispersed CELLs from growing cultures were plated into  siliconized petri dishes. Under these conditions the CELLs spontaneously produced embryoid bodies. Embryoid bodies were cultured in a medium containing retinoic acid for four days and anather four days in a medium without retinoic acid. Then embryoid bodies were plated into tissue culture grade Petri dishes and evaluated morphologicaly. Also cresyl violet staining and immunocytochemical evaluation using nestin and synaptophysin antibodies were performed to verify the phenotype of the neural CELLs. Results:Our results showed after transfering of embryoid bodies into tissue culture dishes neural like CELLs were appeared at the margin of embryoid bodies. Gradually these CELLs increase in namber and grow on the surface. The results of cresyl violet staining and immunocytochemical evaluations confirmed that differentiated CELLs were of neural origin. Conclusion: Therefore, It can be concluded when ever embryoid bodies are  prepared from CCE embryonic stem CELLs using siliconized petri dishes, they can be differentiated into neural CELLs, and CCE embryonic stem CELLs is a suitable CELL LINE for in vitro studies on embryonic development.

Aim: The aim of this study is to examine the efficiency of DNA absorption and gene transfusion to chicken hepatoma CELLs LINE LMH at both transfection and transduction levels using recombinant lentiviruses. Then transgene expression level was studied.Material and Methods: The CELLs were cultivated in 96-well plates for viral transfection or transduction and then studied using fluorescent microscope in different stages. Green Fluorescent Protein-positive (GFP) CELLs counted by Grid CELL Counter software and transgen expression level calculated using ImageJ software. Data analyzed using SPSS.Results: GFP gene expression began 6 and 9 hours after transfection in HEK and LMH CELLs, respectively. These times were increased, respectively, to 22 and 36 hours post-transduction. Counting GFP+ CELLs 48 hours post-transfection showed that HEK and LMH CELLs have received and expressed 40% and 35% of DNA. These levels increased to 95% and 48% respectively in 72 hours. Also counting HEK+ CELLs at 72 and 96 hours viral post-transduction confirmed the similar pattern of receiving DNA by both HEK and LMH CELLs. Therefore, analyzing of GFP expression level showed that HEK and LMH classes expressed GFP in 74% and 89% (P<0.01 significant).Conclusion: Overall, our results indicated that HEK CELLs have higher potential to absorption of foreign genes whereas the intensity of expression is higher in LMH CELLs.

Aim and Background: Cancer is a major cause of mortality in the world. Treatment of Cancer through drug therapy/chemotherapy faces the challenges of drug delivery. Drug delivery systems such as niosomes provided high efficiency and targeted treatment. The aim of this study was to prepare a new formula of niosomal nano system containing doxorubicin as the cancer therapy drug. Changing the tissue distribution of drug leads to drug effect improvement and reducing its cytotoxicity. Materials and Methods: Doxorubicin encapsulated niosomes were synthesized using thin-film method. A certain amount of Span-60 and Cholesterol dissolved by chloroform in a round bottom balloon. Rotary evaporator was used in order to remove the organic solvent and form the thin-film. The thin-film was hydrated by doxorubicin and PBS solution using rotary evaporator in 10° C higher than span-60 transition phase. Entrapment Efficiency and release profile were investigated using dialysis and spectrophotometry. Size reduction was applied by probe sonicating. Nanoparticles average diameter and zeta potentiol was measured using DLS technique and Zeta sizer. The optimum formula was PEGylated and its cytotoxicity investigated through MTT assay on Ovarian cancer CELL LINE and fibroblast normal CELLs. Results: The diameter of optimized and PEGylated niosomal formulation was 172. 5 nm and entrapment efficiency of PEGylated doxorubicin system was about 95. 83± 1. 18. release studies result in PBS buffer at 37° C was the sign of PEGylated nanoniosomes efficiency in controlling the drug release, which the release percent of drug after 48 hours was 20. 52 and its cytotoxicity was higher than the free drug. Conclusion: The study demonstrates using of drug delivery carriers such as synthesized nanoniosomes has an effective role in increasing the efficacy and reducing the consumed dosages of drug.

Background: Hyperglycemia is one of the important featurs of diabetes. In CELL culture studies different methods are used to mimic the hyperglycemia condition. In this study we investigate response of human liver cancer CELL LINE (HepG2) to high insulin, high glucose, and high insulin/ high glucose medium exposure. Methods: HepG2 CELLs were settled in DMEM+0. 1% FBS or DMEM free-serum medium with high concentrations of d-glucose (30 mm) and/or insulin (1μ M) for 24h after an overnight starving in serum-free medium. The level of hyperglycemia was estimated in the supernatants via GOD-POD method. Results: Serum-free madium with high insulin/ high glucose consentration made the higher level of hypreglycemia in HepG2 CELLs. Conclusions: Our study interduced high insulin/ high glucose treatment as the best way to induction hyperglycemia.