مجله سلول و بافت
سال هشتم شماره 3 (پاییز 1396)

Aim: Investigating the effect of catechin hydrate (CH) and boric acid (BA) on rat bone marrow mesenchymal stem cells (MSCs).
material and MSCs were treated with culture media containing CH, then with the help of trypan blue the viability was investigated at 12, 24 and 36hours. 400 and 3200µM of CH along with 6ng/ml of BA and 36hours were selected for further study. Proliferation based on colony forming assay (CFA) and population doubling number (PDN), morphology, level of sodium, potassium and calcium and activity of LDH,ALP,AST,ALT were analyzed. Malondialdehyde (MDA), total antioxidant and activity of SOD and CAT were measured too.
only 3200µM at 12hours and from 400 to 6400µM at 24 and 36 hours, the CH caused significant reduction in viability, proliferation, nuclei diameter and cytoplasm area. Treatment with CH caused increase in activity of LDH,ALP,AST,ALT and increased in FRAP as well as reduction of MDA and sodium, potassium level. BA did not show any effect on viability, morphology and PDN but caused reduction of CFA and activity of LDH,AST and ALT. BA also caused elevation of ALP activity and level of calcium, sodium, potassium as well as MDA level and activity of CAT and SOD. Co-treatment compensated the viability, proliferation, morphological changes, metabolic enzyme activity variation and level of electrolyte to some extent. On the other hand, co-treatment showed, CH ameliorated the oxidative stress induced by BA.
since boron ameliorated the CH toxicity, along with tea we may consume dry grapes and dates.

Aim: The purpose of this research was to design and prepare a suitable gene construct and transfer it to the tobacco plant and analysis of transgenic plants.
Material and Seed-specific construct that containing Napin promoter, Ω sequence, GUS gene, and SAR sequence was prepared in pBI121 plasmid and proliferated in E.coli.. Then tobacco leaf explants were inoculated with LBA4404 agrobacterium strain by standard protocol. Selection of regenerated shoots also were performed in a selection media (co-culture medias 25 mg/L Kan 200 mg/L Cef). Transgenic plants were analyzed by PCR, RT-PCR and histochemical assay.
Analysis of regenerated plantlets by using PCR and specific primers of nptII and GUS indicated that transfer of these genes to plantlets was successful. RT-PCR reaction results showed that nptII is transcribed in both tissues while GUS is transcribed only in the seed tissue. This finding was expected because Nos promoter (which controls the nptII transcription) is a constitutive and Napin is a seed specific promoter.. Expression and activity of Beta-glucuronidase enzyme in seeds of selected plants was confirmed by SDS-PAGE and histochemical assay.
The result of this research showed that designed gene construct was appropriate, because Napin promotes the expression of GUS gene in the seeds and Omega sequences have also been effective in increasing of transgene expression. In the fallowing, this construct can be used for the production of recombinant proteins by replacing valuable genes with GUS..

Aim: Fabrication of biocompatible single and composite nanofiber scaffolds having less interactions in vivo (lack of immune response in the body) and providing optimal conditions for the growth and proliferation of cells.
Material and First, the electrospinning method was used to fabricate three samples of single polycaprolactone (PCL), single polyurethane (PU) and composite PCL/PU (50/50) biopolymer nanofibers. The samples were sterilized with ethylene oxide for 14 hours and implanted under the skin of 12 rats. Also, to examine the growth and proliferation of cells, produced structures were evaluated by the cytotoxicity test (MTT Assay).
Although the rats did not receive antibiotics, localized and systemic reactions and infections of the surgical sites were not observed until the end of the study period. Use of single PCL biopolymer caused negligible edema, fibrosis, and a mild foreign body granuloma. While the PU sample caused the fewest side effects: mainly a negligible edema and moderate fibrosis were observed, but no foreign body granuloma was shown. Also, the PU/PCL sample caused a moderate edema and foreign body granuloma and a mild fibrosis.
Single and composite structures of PCL and PU were all suitable environments for the cell growth and proliferation, as well as the reaction of the immune system in these samples was very low and at a satisfactory level.

Aim: Cancer is a life threatening disease determined by uncontrolled differentiation and proliferation of cells. Chemotherapy is the conventional method in treatment of cancer, but the tumor often will be resistant to chemotherapy regimen leading to crucial side effects. Many bio-toxins are biologically active compounds with anti-tumor activity. This study was aimed to find an anti-cancer agent from the venom of Iranian cobra snake.
Material and The process of collection, lyophilisation of Iranian cobra (Naja naja oxiana) venom. Protein concentration of Iranian cobra venom was determined by BCA assay. Fast protein liquid chromatography (FPLC), SDS-PAGE, Anion exchange chromatography were used for purification of Iranian cobra venom. Standard cell biology methods were employed to characterize Iranian cobra venom abilities (in vitro) to inhibit Platelet aggregation, adhesion, migration and invasion of tumor cells. Its anti-cancer activity in (Jurkat E6.1) (in vitro) was tested by MTT assay.
Peak 3 of FPLC was active on cell lines and selected for anion exchange chromatography. Four anionic and one cationic fractions collected for anticancer activity. Peak 2 of anion exchange chromatography had the most toxic activity as 43%±1 at 1.5 micrograms. This fraction showed about 8%±1 toxicity on fibroblast cells. (P-value ≤ 0.05)
Natural anti-cancer compounds derived from the Naja naja oxiana venom, can fight against cancer. It is the first report of an anticancer fraction from Iranian cobra with toxicity on Jurkat E6.1 cell line with less toxicity to normal cells.

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 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.
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.
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.

Aim: The present study aims to investigate the effect of simultaneous encapsulation of DNA with different ratio of PLL via PLA-PEG copolymer, on gene delivery efficiency into mammalian cells. Some characteristics such as biocompatibility, DNA protecting against restriction enzymes, DNA release rate, size and zeta potential were also investigated.
Material and PLA-PEG/PLL/DNA nanoparticles with different ratio of PLL were prepared by double emulsion-solvent evaporation technique. Then, the release percentage of DNA and the zeta potential of particles in Phosphate Saline buffer (PH=7) were measured.
Increasing the PLL percentage in the PLA-PEG/PLL/DNA nanoparticles resulted in enhancement of particles size and zeta potential. Gel electrophoresis analysis of DNA extracted from PLA-PEG/DNA and PLA-PEG/PLL/DNA nanoparticles after treatment with DNase I enzyme indicates the ability of PLA-PEG and PLA-PEG/PLL copolymers in DNA protection. The flow cytometry and fluorescent microscopy study confirmed that gene transfer efficiency was improved by increasing the ratio of PLL in PLA-PEG/PLL/DNA nanoparticles. Moreover we found that gene transfer efficiency was one and half fold higher then PLL/DNA complex in the serum medium.
The results showed that PLA-PEG nanoparticles in the serum medium have higher potential gene delivery to the MCF-7 cells in comparison with PLA-PEG/PLL/DNA nanoparticles.