Cell and Tissue Journal

Abstract Aim: The current study was designed to understand the destructive effects of mancozeb on BTB in an in vitro model.
Material and Methods: Male mice were gavage- administered MZB (250, 500 mg/kg w. w) for 40 consecutive days. Integrity of BTB was evaluated using Evans blue dye. The relative mRNA expression of BTB- associated genes; including N- cadherin, claudin-11 and zonula occludens-1 was investigated by quantitative real-time reverse transcription–polymerase chain reaction in different groups (qRT-PCR).
Results: Concentration of Evans Blue dye in the lumen of seminiferous tubules was increased in mancozeb-treated animals. The mRNA expression levels of N-cadherin, claudin-11 and zonula occludens-1 genes were decreased significantly in the MZB 500 mg/kg group compared with the control group.
Conclusion: According to the obtained data, it could be concluded that mancozeb increase BTB permeability leading to testicular dysfunction by the alteration of BTB-associated genes.
 

Abstract Aim: In this study, we use combination of enzymatic and explant methods, for isolation and culture of ADMSCs.
Materials and Method: Adipose tissues dissected out from the abdominal region of male Wistar rats. Tissues were minced into small pieces (1-2 mm), and then treated with trypsin-EDTA 0.25% for 30 minutes at 37 ̊C in shaker-incubator. Enzymatic treated tissues were centrifuged and floating parts were cultured. Statistical analysis of the cells number was investigated using GraphPad Prismv6 software.
Results: Results showed that ADMSCs isolated with our methods have growth characteristics similar to conventional methods. ADMSCs were maintained up to 10th passage and exhibited a homogeneous population in terms of appearance and morphology. We demonstrated that ADMSCs by these combination methods have mesenchymal characteristic markers (positive for CD90, CD44, CD73, CD105 and negative for CD35, CD45, CD11b).
Conclusion: In the current study, we showed that the combination of enzymatic and explant methods creates a simple, inexpensive and reproducible method for isolation and culture of ADMSCs.
 

Abstract Aim: The aim of this study was study the role of Resveratrol (RSV) on proliferation and colony efficiency of dissociated human embryonic stem cells (hESCs).
Material and methods: In the current study, we used HESC line, RH6. Cells were cultured on the matrigelin supplemented hESC specific medium. Cell proliferation was estimated using cell counting and Brdu incorporation assays. The expression of colony efficiency markers (E-cadherin and β-catenin) was evaluated by western blot technique and β-catenin localization examined by immune-cytochemistry staining.
Results: We have shown that RSV promoted hPSCs proliferation without affecting their colony efficiency.
Conclusion: According to these observations, RSVcan be suggested as a new supplement for hESCs culture.
 
 

Abstract Aim: In current study, we aimed to reduce Cdc42 gene expression in lung carcinoma related cells, Calu-6, and assayed its effect on cell proliferation.
 Material and Methods: To reduce the expression of Cdc42 gene shRNA system was used and lentiviral system was selected to deliver Cdc42 specific shRNA to Calu-6 cells. Recombinant lentiviruses produced by co-transfection of pMD2G, psPAX2 and p-GFP-C-shLenti plasmids into 293T cells using lipofectamin. Efficiency of transfection and transduction assessed by florescent microscopy. Viability of cells treated by recombinant lentiviruses assessed by MTT assay.
 Results: florescent microscopy showed 80% transfection of 293T cells and high rate of Calu-6 cells transduction. MTT assay results revealed that viability of transduced Calu-6 cells reached to %58 and %40 in compare to control and negative control cells, respectively.
Conclusion: recombinant lentiviruses properly transfer Cdc42-shRNA into Calu-6 cells, leading to reduction of cell proliferation. Silencing of Cdc42 gene expression using lentiviruses is persist and long-term effect which can be under attention for gene therapy of lung cancer.
 

Abstract Aim: This investigation was conducted to determine the amount of isolated anthocyanins from seed coats of Phaseolus vulgaris L. under magnetic field treatment and also assess their antiproliferative activity of the extracts.
Material and method: Dry and soaked red bean seeds were subjected to different electromagnetic field treatments in four group. The seeds of groups 1(dry) and 2 (soaked) were treated for 45 min., while in groups 3 and 4 the dry and soaked seeds treated two times with 120 min. interval with electromagnetic field.   Then, they were cultured and grown, and their produced seeds were collected. Seed coats from these beans were separated and coat extract was prepared. Two groups of anthocyanins, cyanidin and pelargonidin, were identified and measured by high performance liquid chromatography (HPLC). The antiproliferative activity of the extract was determined by MTT assay against ovarian cancer cells (CP2780A).
Results: The bean coats of the control and the treated plants contain a high content of cyanidin and pelargonidin. Among the extracts, the samples of group 1 had the highest amount of cyanidin, while seeds of group 2 had the highest amount of pelargonidin. The results of the antiproliferative activity revealed that the methanol extract of the treated red beans under 4 mT were highly antiproliferative activity of the cell (78.74 to 84.44 %). Moreover, the seeds of group 2 showed the highest antiproliferative activity (IC₅₀: 72.63 ± 2.2).
Conclusion: The electromagnetic field treatment is the most effective in increasing the amounts of anthocyanins in red beans, and that the red bean coat can be used as a natural source of nutrition and drug supplement with antiproliferative properties.

Abstract Aim: Aquaporin4 (AQP4) is the main water channel in the brain that has important role in water transport across blood- brain- barrier. It has been proved that intracranial pressure and also expression of AQP4 increase in some disorders such as Hydrocephalus, Hyponatremia, cytotoxic oedema and brain tumours and drugs that can decrease the expression of this AQP can offer some treatment for these disorders. The aim of this study was to determine whether green tea extract (GTE) can down regulate the Aquaporin4 protein level in brain cortex.
Material and methods: This research is a laboratory experimental study. 40 Wister rats (4-6 weeks) were randomly divided into 4 groups as follows: untreated control group, sham group treated intra peritoneally (IP) with 200 µl saline (solvent GTE), experimental1&2treated IP with 100& 200 mg/kg GTE dissolved in 200 µl saline. After 6 hours, expression of AQP4 in control, sham, and experimental groups were tested by immunohistochemistry and western blotting.
Result: Results from immunohistochemistry and western blotting showed that GTE can down regulate the AQP4 expression level on dose-dependent manner in brain cortex.
Conclusion: It is suggested that GTE has potential to reduce the level of AQP4 protein in brain cortex and it can be useful as a herbal medicine to reduce intracranial pressure in diseases such as hyponatremia, cytotoxic oedema, brain tumours and etc.

Abstract Aim: In this study the effect of vitreous humor on the mesenchymal stem cells (MSCs) derived from bone marrow, adipose tissue and amniotic fluid to lens like cells was investigated.
Material and methods: Inthis experimental study collected stem cells from femurs bone marrow and inguinal fat pads of mature NMRI mice and amniotic fluid of 13 days embryos of NMRI mice were cultured. The mesenchymal character of these cells was proven by flow cytometry markers like CD90, CD31. This experiment took place for 14 days with different dosages 10, 15, 20, 30, 40, 50 % of bovine vitreous humor. Express of crystalline markers were detected by immunocytochemistry in experimental and control groups.
Results: The flow cytometeric analyses of surface markers were shown expression of CD90 by bone marrow and amniotic fluid stem cells and adipose tissue stem cells (29/29%, 0/57%, 82%, respectively). Moreover, mesenchymal stem cells from these 3 sources had expression of CD31 (3/44%, 1/53%, 0/1%, respectively). Immunocytochemistry  results revealed that 40 % vitreous humor in culture media fluid more inducing effect on adipose and amniotic fluid mesenchymal stem cells and 15 % vitreous humor on bone marrow stem cells in order to differentiate.
Conclusion: According to the findings of this study, it can be concluded that MSCs derived from all sources can differentiate into lens fiber like cells by inducing effect of vitreous humor.

Abstract Aim: The aim of present study was to demonstrate that the obstruction of CSF flow in hydrocephalic Texas (HT-x) rat and the resulting changes in its protein components can cause abnormal cortical development.
Material and Methods: Cortical cells were taken from E18 and E21 Wistar fetuses and cultured for 24 hours in Neurobasal medium. Two groups of experiments were explored. These cells were treated with heated and non-heated CSF collected from E20 and E21fetuses. The cells were cultured for a further 48 hours.At the end of this time, proliferation assays and morphological analysis were performed
Results: Prenatal affected CSF had an inhibitory effect on the proliferation of normal primary fetal cortical cells. Heating the CSF removed this effect.
Conclusion: CSF contains important factors that can stimulate both proliferation and cell differentiation and that some of these are likely to be proteins. Disruption of CSF flow can cause the cortical deficiencies in patients with hydrocephalous.