Cell Journal (Yakhteh)
Year:2007 | Volume:8 | Issue:4 (32)|Papers Count:8

Introduction: Determination of best culture medium for fertilization and development of NMRI mice.Material and Methods: Development of fertilized mature oocytes of NMRI mice was studied for five days in several culture media (KSOM, T6, M16, CZB, G1TM ver3 and G2TM ver3) following hormone therapy with PMSG and HCG.Results: The rate of fertilization after 24 hours was 97.4% in KSOM, 83.5% in T6, 90.5% in M16, 100% in CZB and 90.9% in G1TM ver3 and G2TM ver3. The differences between CZB and KSOM culture media in comparison to T6 and M16 and G1TM ver3 and G2TM ver3 media were statistically significant (p<0.05). Blastocyte stage was seen in 96.3% of CZB group which was the highest in comparison to other culture media (p£0.05). Blastocyte formation was 80.2% in T6, 82.1% in M16, 77.6% in KSOM and 87.5% in G1TM ver3 and G2TM ver3 culture media. The highest Hatching blastocyst rate was seen in sequential medium (G1TM ver3 and G2TM ver3) and KSOM culture medium (76.1% and 66.4%, respectively) while the lowest rate was 60.3% in M16 medium. The rate of embryo degeneration was lowest in sequential medium (G1TM ver3 and G2TM ver3) (23.9%) in comparison to other groups and the difference was statistically significant regarding T6 group (51.6%) (p<0.01)Conclusion: This study reveals that CZB, KSOM and G1TM ver3 and G2TM ver3 are the best culture media for pre implantation embryo development of NMRI mice. The difference in development of mice embryos in different culture media can be due to difference in medium supplements (aminoacids) and concentration of culture medium components.

Introduction: Study the protective effect of mucosal BCG vaccination with subcutaneous against Leishmania major infection in susceptible BALB/c miceMaterial and Methods: 6-8 weeks old Male BALB/c mice were divided to two test groups and four control groups of 20 of mice and then vaccinated. Two test groups vaccinated with appropriate dose of BCG through rectal and subcutaneous routs. They vaccinated one month later with ALM+alum subcutaneously. After receiving the booster dose (21 days later), mice challenged with the injection of 105 leishmania major in the hind footpad and lesion development evaluated every week. Immunologic responses like, delayed type hypersensitivity to Purified Protein Derivatives (PPD), splenocytes proliferation and IFN-g and IL-5 cytokine production measured in the supernatant of splenocytes culture. Control groups were those that received only ALM+alum or only alum adjuvant subcutaneously. Another control groups challenged with Leishmania without any vaccination or were kept as environmental control without any treatment.Results: stimulatory effect of BCG on immune system observed in both groups of vaccination according to significant increase (p£0.05) in delayed type hypersensitivity and IFN-g production response. Increase in splenocytes proliferation observed up to three weeks after challenge. But after that, BCG vaccinated groups had a lot of variations in immune response that finally tend to disease overcome in subcutaneously BCG vaccinated group. The main reason was the significant decrease in IFN-g production and splenocytes proliferation to rectal group .Parasite dissemination in lymphoid organs were increased significantly in comparison with rectal immunized group. In a different way rectal immunized group showed a high resistant to infection by increase and maintenance of anti-leishmania responses.Conclusion: Rectal BCG vaccination can induce more stable cellular immune response in comparison to subcutaneous vaccination and make animals protective, by inhibition of L. major dissemination.

Introduction: There are some evidences that the Matricaria recutita extract has sedative effects and relieves symptoms of morphine withdrawal syndrome. Symptoms of this syndrome are due to increased expression of fos transcription factor in brain. In this study the effect of chronic hydro alcoholic extract (30mg/kg) of Matricaria recutita on jumping behavior and expression of fos transcription factor during morphine withdrawal were investigated in male mice.Material and Methods: Mice were divided into 5 groups (morphine + saline, morphine + naloxone, morphine + chronic extract of Matricaria recutita + naloxone, morphine + saline + naloxone, saline + naloxone). To develop morphine dependence, mice were injected subcutaneously with increasing doses of morphine for 4 days. Mice received a final morphine injection (40mg/kg) 3h prior to naloxone (5mg/kg) on the fourth day of testing. For chronic application, Matricaria recutita extract (30mg/kg, intra peritoneal) was administered concurrently with morphine for 4 days. To study the behavior of mice, jumping sign of morphine withdrawal was assessed 30min after naloxone injection. In cellular study, 90min after the naloxone injection, mice were decapitated and their brains were separated, then mRNA was extracted from brain tissues. Using DIG-labeled DNA probe of C-fos and beta-actin and dot blot technique, expression of C-fos was analyzed by Zero D scan software.Results: The rate of jumping sign due to abstinence from morphine was increased compared to control groups and chronic application of Matricaria recutita extract at 30mg/kg led to significant decreases in jumping sign and amount of expression of C-fos after naloxone-precipitated abstinence.Conclusion: It seems that the sedative effect of Matricaria recutita extract is via modulation of c-fos expression; therefore it has a permanent inhibitory effect regarding morphine dependence.

Introduction: Different microbial compounds can induce maturation of dendritic cells (DCs) into DC1 or DC2 .In the present study, we used tumor antigen pulsed dendritic cell that matured with Listeria monocytogenes antigen and cholera toxin (CT) in the treatment of a murin model of cancer.Material and Methods: Tumor cells (WEHI-164 BALB/C derived fibrosarsoma) were injected subcutaneously to Balb/c mice. Bone marrow cells were cultured with GM-CSF and IL-4.On day 5, Listeria monocytogen lysate or CT (cholera toxin) added to culture for 2 days.For immunization, 106 dendritic cells matured with CT or Listeria monocytogenes (L.m injected) subcutaneously around the tumor site.Results: We found that Immunotherapy with dendritic cells treated with listeria antigens leads to delay in tumor growth as compared to the CT and control groups. DCs treated with Listeria produce higher amount of IL-12 compare to CT and control groups.Conclusion: Some microbial components such as Listeria monocytogenes that favors the immune response towards TH1 could be used in generating powerful DCs for cancer Immunotherapy.

Introduction: Human calprotectin is an abundant protein in the neutrophil, monocyte and macrophage cytosol. Effect of proliferation inhibition of this protein has been elucidated on hepatoma, melanoma, murine fibrosarcoma lukemia and human adenocarcenoma breast cancer but not in human gastric cancer cell lines. In present study, in vitro proliferation inhibition effect of calprotectin in different concentrations and intervals on AGS (tumoral) and HGF (normal) cell lines was evaluated.Material and Methods: Calprotectin was purified from human neutrophil by chromatography methods. The human gastric adenocarcinoma cell line (AGS) was used. These cells were maintained in RPMI 1640 medium supplemented with 10% FCS. AGS cells (10000 cell per well) were exposed to different concentration of calprotectin (25, 50, 100, 200, 400 µg/ml) at 24, 48, 72 h. For evaluation of calprotectin on normal cells, human gingival fibroblast (HGF) was incubated with 70µg/ml of calprotectin (almost 2 time of LC50 evaluated in AGS after 48 h) in this study MTT assay was used for evaluation of proliferation inhibition effect of calprotectin on AGS cells.Results: Viability of the cells with increasing of calprotectin concentration was decreased. All of the cells in 400 µg/ml of calprotectin after 24 h were dead. Although, toxicity of the cells with increasing of incubation time was increased The most cytotoxicity effect was evaluated on 72 h .The LC50 of calprotectin for AGS cell at 24, 48 and 72 h, relief 96.78, 38.66 and 9.86 µg/ml  was determined, respectively. Comparison between proliferation inhibition effect of calprotectin on AGS cell and HGF was showed that difference is not significant but more effect on AGS cell lines.Conclusion: These results showed that in compare to normal cell (HGF), calprotectin has more cytotoxicity effect on cancer cells (AGS). Calprotectin can be candidate as an anti-gastric cancer drug.

Introduction: Endometrial receptivity towards embryo implantation involves several dynamic changes, i.e. “the plasma membrane transformation of the luminal epithelium”. This includes the appearance of large projections on the apical surface of the epithelial cells, which their function(s) has not been clearly known in humans. This study is designed to identify the potential physiological roles of these projections (pinopods or uterodomes) during the human embryo implantation.Material and Methods: Endometrial biopsies from early, mid-, and late luteal phases of the menstrual cycle of 23 fertile female patients with regular menses were taken. To study the physiological function of human pinopods as biological markers of endometrial receptivity, scanning and transmission electron microscopies (SEM and TEM) as well as immunofluorescent and immunogold TEM were performed. Double- staining of light and electron microscopy were performed to detect leukemia inhibitory factor (LIF) and sintaxin-1, vesicle–associated membrane protein-2 (VAMP-2), and 25 kDa synaptosomal protein (SNAP-25) in order to identify their special correlation with human pinopods.Results: Pinopods (uterodomes) were abundant in the human endometrial glands. Morphological studies suggested secretory function for both the luminal and the glandular uterodomes. Images of immunostanings revealed co-localization of LIF with the specific markers of exocytosis.Conclusion: Uterodomes of human endometrium have secretory function, which is far from the pinocytotic role of pinopods of rodents.

The increasing prevalence of chronic diseases, e.g., cardiovascular disease, diabetes and neuronal degenerative disease, present a challenge to find more effective therapies. Stem cell-based therapy, including embryonic, umbilical cord blood and adult stem cells provides a rational treatment tool for regenerative medicine and has the potential to revolutionize modern therapy. However, there are many obstacles to the successful outcome of transplanted stem cells. One of the most important obstacles which has limited their practical usefulness is immunological barrier. This review will focus on the potential mechanisms that the immune system could use to target transplanted stem cells and how unwanted responses might be prevented.

Cell Banking is long-term storing of cell stocks that requires intensive care and maintenance to ensure its adequate quality.The main goal of a biological bank is preparing a repository source of cell culture system in a global manner. On the other hand, cell culture is an invaluable tool for investigators in numerous fields. Successful maintenance of cells requires knowledge and practice of some essential techniques. Many areas of medical research and therapeutic programs have benefited from the use of well characterized cell lines and tissue bankable cells. According to above mentioned, initiating material and safety aspects of handling cell lines is most important in controlling freezing rate.Through another section of this article, extensive discussion about mechanisms of biological characterization of cell population is defined. Some of the techniques about in vitro manipulation for complete processing of banked cells for improving proliferative capacities, culture longevity, and stability in various source of cells have been discussed in terms of quality control and banking procedures. For highly standardized procedures we recommended some quality control of cell line identity and methods for eradication of mycoplasma contamination and other microbial agents, since they have multitude effects on cell banks.