Vildagliptin enhances differentiation of insulin producing cells from adipose-derived mesenchymal stem cells

Objective: Type 1 diabetes is caused by destruction of beta cells of pancreas. Vildagliptin [VG], a dipeptidyl peptidase IV [DPP IV] inhibitor, is an anti-diabetic drug, which increases beta cell mass. In the present study, the effects of VG on generation of insulin-producing cells [IPCs] from adipose-derived mesenchymal stem cells [ASCs] is investigated

Materials and Methods: In this experimental study, ASCs were isolated and after characterization were exposed to differentiation media with or without VG. The presence of IPCs was confirmed by morphological analysis and gene expression [Pdx-1, Glut-2 and Insulin]. Newport Green staining was used to determine insulin-positive cells. Insulin secretion under different concentrations of glucose was measured using radioimmunoassay method

Results: In the presence of VG the morphology of differentiated cells was similar to the pancreatic islet cells. Expression of Pdx-1, Glut-2 and Insulin genes in VG-treated cells was significantly higher than the cells exposed to induction media only. Insulin release from VG-treated ASCs showed a nearly 3.6 fold [P<0.05] increase when exposed to a highglucose medium in comparison to untreated ASCs. The percentage of insulin-positive cells in the VG-treated cells was approximately 2.9-fold higher than the untreated ASCs

Conclusion: The present study has demonstrated that VG elevates differentiation of ASCs into IPCs. Improvement of this protocol may be used in cell therapy in diabetic patients

effect of the melatonin on cryopreserved mouse testicular cells

Background: After improvements in various cancer treatments, life expectancy has been raised, but success in treatment causes loss of fertility in many of the survived young men. Cryopreservation of immature testicular tissues or cells introduced as the only way to preserve fertility. However, freezing has some harmful effects. Melatonin, a pineal gland hormone, has receptors in reproductive systems of different species. It is assumed that melatonin has free radical scavenger properties

Objective:The aim of this study was to evaluate the effects of melatonin on the cryopreserved testicular cells in mouse

Materials and Methods: Cells from 7- 10 days old NMRI mice testes were isolated using two step enzymatic digestion. The testicular cells were divided into two groups randomly and cryopreserved in two different freezing media with and without the addition of 100 micro m melatonin. Finally, apoptosis of the cells was assayed by flow cytometry. Also, lactate dehydrogenase activity test was performed to assess the cytotoxicity

Results: The results of lactate dehydrogenase showed the nearly cytotoxic effect of melatonin. The results of flow cytometry showed increase in apoptosis in the cryopreserved cells in the media containing melatonin compared to the control group

Conclusion: The present study shows that melatonin has an apoptotic effect on cryopreserved mouse testicular cells

Immunohistochemical assessment of galectin-3 during pre-implantation in mouse endometrium

Background: galectin-3 [Gal-3], a beta-galactoside-binding lectin, is a multifunctional lectin that involves in a number of critical biological processes

Objective: the purpose of this study was to investigate the expression pattern of Gal-3 in mouse endometrium during estrus phase of estrous cycle and pre-implantation

Materials and Methods: in this experimental study 42 NMRI female mice were divided in seven different groups. Ovulation in NMRI female mice was stimulated by injecting hMG and hCG. Estrus phase was considered as stimulated and un-stimulated groups. The other groups of mice were mated, and the day of vaginal plug formation was considered as the day 1 of pregnancy. The mice of all groups were sacrificed on different days of pre-implantation period and their uterine horns were fixed and avid in- biotin complex method of immunohistochemistry [IHC] was applied

Results: in estrus group, Gal-3 immunoreactivity in luminal epithelium was strong, in stromal cells very strong, in glandular epithelium very weak and endothelial cells very strong. No identifiable difference was observed in un-stimulated and stimulated estrus phase. In test groups, days 1-2, insignificant difference of Gal-3 expression was observed. On day 3, luminal epithelium and stromal cells showed significant decrease in comparison to estrus and day 1 [p=0.001]. On the 4[th] and 5[th] days, luminal epithelium and stromal cells showed significant decrease in comparison to estrus phase and days 1-3 [p=0.0001]

Conclusion: the data suggested that successful implantation is probably associated with the downregulation of Gal-3 in the mouse endometrium at the beginning of pregnancy

[ comparison of the effect of honey and vitamin E on sex hormone levels in male wistar rats exposed to noise pollution]

Noise pollution has a high prevalence among the environmental pollutions and is considered as a teratogenic agent for reproductive system. This study was performed with the purpose of evaluation of the effect of honey and vitamin E on sex hormone levels in male rats exposed to noise pollution. Twenty-four adult male rats with the mean weight of 200 +/- 20g were randomly divided into 4 groups: 1 [honey+voice], 2 [vitamin E+voice], 3 [voice], and 4 [control]. Groups 1 and 2 received honey and vitamin E as gavage, in addition to voice; group 3 was only exposed to noise pollution. After 50 days, serum level of hormone in male rats were measured by ELISA technique after taking blood from heart. Then, the male rats were placed in a cage with female rats of the same breed with a 2:1 ratio. Weight and number of embryos from fertilization were assessed. Data were analyzed using ANOVA and Tukey statistical methods. In this study, it was found that the secretion of sex hormones [FSH, LH, and testosterone] impaired under the effect of noise pollution. Serum level of testosterone decreased in rats that were under noise stress [p<0.05], and use of honey and vitamin E as antioxidants were modulated the level of this hormone in male rats. Noise pollution in male rats increased in the serum level FSH, LH [p<0.05]. Weight and number of live embryos decreased because of this stress [p<0.05]. The use of Honey and vitamin E by male rats increased live embryos [p<0.05]. According to the results of this study, noise pollution has negative effects on the fertility of male rats. Also, use of honey and vitamin E increases the fertility in groups exposed to noise pollution

in vitro fertilization rate of mouse ova in the absence or presence of recombinant human leukemia inhibitory factor

To examine the effect of human recombinant leukemia inhibitory factor in different doses on rate of fertilization of mouse ova. Prospective study. Department of Anatomy, laboratory of cell culture. Animals: Female NMRI mice 6 to 8 weeks old. Mice were killed at 12-14 hours after hCG or 36-38 hours after hMG injection. Mature oocytes were obtained and divided randomly into 5 groups. Oocytes in group A [n=157] were cultured as the control group in TYH medium. Oocytes in groups B, C, D, E [n=137, 154, 166 and 159, respectively] were cultured in the same medium supplemented with recombinant human leukemia inhibitory factor in four different concentrations [5, 7.5, 10, 20ng/ml, respectively] for 1 hour. After that time 100000 spermatozoa were added to every drop and after 24-26 hours two cell embryos were recorded. Fertilization was assessed by recording the number of 2-cell embryos and analysed by X[2] tests. Two cell embryos. No significant difference was detected in the rate of two cell embryos in the studied experimental groups as compared with the rate of two cell embryos in control group [Group A]. This study showed that, different concentrations of recombinant human of leukemia inhibitory factor in standard medium does not enhance in vitro fertilization rate of mouse oocytes

Vitrification of human oocyte using cryoloop

The cryopreservation of human oocyte would make a significant contribution to infertility treatment, such as using it for oocyte donation and for patients a bout to lose ovarian function due to surgery or chemotherapy. Despite of using st and ard freezing straws and cryovials or even open pulled straws, only a few successful pregnancies have been arisen from cryopreserved human oocytes. This situation has been primarily attributed to poor survival, fertilization and development of cryopreserved oocytes. The aim of this study was to evaluate the novel cryoloop vitrification method for cryopreservation of human oocytes. Nine infertile couples participated in this study. In all women proper regulation and desensitization was done using GnRH agonist during luteal phase. Mature oocytes allocated into two groups r and omly. In group I, 34 oocytes were vitrified in conventional straws, while in group II, 33 oocytes were vitrified in cryoloop. After a store time of 1-6 months the oocytes were thawed, incubated for 2 hours and subsequently the ICSI was done on survived oocytes. To verify normal fertilization of vitrified oocytes the number of pronuclei in the cytoplasm was counted 16-18 hours after ICSI and good morphological quality embryos were transferred on day 2 or 3 after sperm injection. Pregnancy was identified by the serum beta HCG level, checked 14 days after embryo transfer. The present study shows that the rate of survival of vitrified human oocytes in two groups has no significant difference [52.94% in group I versus 63.63% in group II] but the fertilization rate of vitrified oocytes by cryoloop was greater than vitrified oocytes by conventional straws [73.7% versus 55.55% respectively]. One of the embryo transfers achieved clinical pregnancy and resulted in the delivery of healthy baby. Vitrification by using cryoloop can improved the fertilization rate and developmental capacity of vitrified thawed oocyte