Cell and Tissue Journal

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 Aims: In this study, the effects of Vitamin D3 on total protein concentration (TPC) and MBP expression in the corpus callosum extracts of Cuprizone induced demyelinated mouse has been investigated.
 Material and Methods: The mice were treated by Cuprizone for five weeks in order to induce demyelination. Then, the mice were divided into 3 groups. The first group was injected intraperitoneally (IP) by vitamin D3 in the amount of 5 µg/kg/daily body weight. The second group (SHAM) was injected IP by phosphate buffered saline (PBS) and the third group was left without injection as controls. After five weeks, the mice were killed and the corpus callosum was collected and the total protein concentration (TPC) and expression of MBP were studied by Western blot technique.
Results: No significant variation in the cortical TPC was seen in vitamin D3 injected group as compared to either SHAM or controls. We have also shown that the expression of MBP in the corpus callosum extracts of demyelinated mouse was significantly increased in the vitamin D3 injected group as compared to the other groups.
Conclusions: It is concluded that vitamin D3 increases MBP expression in the corpus callosum of cuprizone-induced demyelination mice. It is also suggested that vitamin D3 may play a role in the process of remyelination by increasing MBP expression in the cortex.
 
 

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 Cryopreservation is the most effective method for long-term maintenance of sperm. There are several procedures for freeze-thawing of semen and each may impose damage on sperm function, viability and finally decreases semen quality and fertility potential. In addition to decreased percentage of sperm viability and motility after freeze-thawing, percentage of DNA damage is also increases due to high level of oxidative stress. To minimize these damages, we need to increase our insights regarding different cryopreservation procedures, cryoprotectant and antioxidant supplements, which can protect sperm membrane during cryopreservation. Therefore, by using these experiments, we can improve the efficiency of these procedures. In this review, we discuss about principles of cryopreservation, types of freeze-thawing methods, advantages and disadvantages of each of these methods, effects of freezing on sperm parameters and clinical outcomes, and finally role of antioxidants in preservation of sperm integrity during freeze-thawing. For this review, all relevant information was collected via databases such as PubMed and Google Scholar during the period of 1966-2017.
 

Abstract Aim: In this study, the genotoxic effect of low frequency electromagnetic wave  that is similar to mobile phones in wavelength, as a main source of producing this waves, in combination with a known aneugen (vinblastine) was investigated on the cultured cells.
Material and Methods: L929 cells were exposed to two forms of continues and discontinues electromagnetic waves with wave length of 900MGh in absence and presence of 1.5 ng/ml of vinblastine for 2 h. Investigation of induced chromosomal abnormalities and cell viability in all treated groups were performed by micronucleus assay on binucleated cells and MTT assay, respectively.
Results: The frequency of micronuclei of cells in continuous and discontinuous electromagnetic fields was significantly higher in comparison with control and cells in turned off electromagnetic field, which represented higher chromosomal aberrations. Co-treatment of the cells with vinblastine did not increase this frequency. Analysis of toxicity showed no decrease in cell viability in all treated groups.
Conclusion: findings represented the ability of electromagnetic field, especially in form of discontinues, in inducing chromosomal abnormality. Also, co-treatment of cells with electromagnetic filed and vinblastine did not elevate the level of chromosome abnormality.
 

Abstract Aim: The aim of this study is investigation of cell toxicity of oxaliplatin on colon cancer cell line and analysis of caspase 3 and 9 apoptotic genes.
Material and methods: In this experimental study, cytotoxicity of oxaliplatin on colon cancer cell line (HT29) was evaluated using MTT method in different concentrations including 100, 50, 25, 12.5, 6.25 and 3.125 µg/mL and the IC50 value was determined. After treatment of HT29 cells with IC50 value, the cells RNA was extracted and converted to cDNA. Subsequently, the expression level of caspase 3 and caspase 9 apoptotic genes comparing to house-keeping gene (β-actin) was measured using Real Time PCR.
Results: Treatment of HT29 cells with various concentrations of oxaliplatin including show that oxaliplatin has the highest cytotoxic effect in 100 µg/mL, which is statistically significant (p < 0.05). In addition to, the IC50 value of oxaliplatin was 6 µg/mL. Moreover, the expression ratio of caspase 3 and 9 genes comparing to β-actin gene in HT29 cells treated with oxaliplatin were up-regulated (2.69±0.72 (p < 0.001), 3.26±0.56 (p < 0.001), respectively).  
Conclusion: According to cell toxicity and apoptosis induction in HT29 cells by oxaliplatin, it can be concluded that this drug is an appropriate choice for treating of colon cancer.
 

Abstract Aim: The purpose of this study was to investigate the effect of adding different levels of zinc (Zn) to semen extender on the sperm parameters after freezing and thawing in rams.
Material and methods: The spermatozoa of five Farahani rams breed (with a mean age of 3 ± 0.5 years and weight of 60 ± 5 kg) were taken twice a week. The semen was collected from the rams by artificial vagina and then the samples were mixed to removing the individualistic effects. The sperm samples (in five replicates) were randomly divided into four groups (zero, 50, 100 and 150 μM of zinc sulfate). Each falcon had the desired diluent that was mixed with Zn and tris and prepared for freezing steps. Samples were stored in semi-milliliter straw and kept with samples under the same conditions and temperatures of 37 °C. Moreover, in these conditions were filled and frozen. Sperm samples were evaluated after thawing for motility, progressive motility, survival (eosin-neogrosin test), sperm membrane integrity (HOST) and sperm morphology (Hancock test).
Results: Results showed no significant difference in the motility percentage and sperm membrane integrity in zinc sulfate environments compared with the control group. The treatment with 50 μm of zinc sulfate showed a higher level of significant (P˂0.05) in the percentage of progressive motility compared with the control group. In sperm viability test, the treatment with 100 μm of zinc sulfate was significantly different from those of 50 and 150 μm zinc sulfate treatment (P˂0.05).
Conclusion: application of zinc sulfate in ram semen extender might be improving some of sperm parameters after the freezing-thawing.
 

Abstract Aim: One of the most important infraspecific variations is differences of tissues and cells in plant that occur between various populations of the same species for adaption with environmental factors. These variations can be consider as a starter for speciation.
Material and methods: In the present study, leaf anatomical traits of four Nepeta heliotropifolia populationswere examine. Three flowering stems were collected from each population, and from each stem a mature intact leaf was elected. Leaves were fixed in fixative and then transferred to ethanol. Hand sections were decolorized, double stained and their anatomical structure were studied using light microscopy. The MVSP and SPSS soft wares were used for statistical analyses. 
Results: In total, twenty two qualitative and quantitative anatomical characteristics were studied. Qualitative features were stable between populations, while ANOVA test showed significant differences for most of the studied variables. In addition, significant correlations were found between morphological characters with each other and ecological factors of habitats. The studied populations clustered separately in UPGMA tree, PCA and PCO plots.
Conclusion: Environmental factors have effects on tissues and cells of plant and create infraspecific variations. They have more effects on quantitative variables compared with qualitative ones. Leaf anatomical variations of populations have no relationship with distances between the populations, but similarity or difference of ecological factors is a very important factor in creating similarity or difference between populations. The pattern of population’s arrangement in the created tree and plots approved it.