Effects of Different Doses of Bone Morphogenetic Protein 4 on Viability and Proliferation Rates of Mouse Embryonic Stem Cells

Objective: In this study, we examined the effect of different doses of bone morphogeneticprotein 4 (BMP4) on CCE mouse embryonic stem cells (ESCs) viability andproliferation rates in order to improve the outcome of induction processes and make asystem with highest viability and proliferation rates for further studies on BMP4 roles inmultiple developmental stages.Materials and Methods: Expression of Oct-4 was studied and confirmed in this cellline immunocytochemically. Also, in order to evaluate the proliferation and viabilityrates in BMP4-treated cells, ESCs were cultured in Dulbecco's Modified Eagle Medium(DMEM) containing different doses of BMP4 (0, 0.01, 0.1, 1, 5, 25, 50 and 100ng/ml).The mean number of whole cells and living cells were considered as proliferation andsurvival rates respectively. Data analysis was done with ANOVA test.Results: The results showed that there were significant differences between the meanpercent of viability between 1ng/ml and 0 ng/ml (control) and 50 and 100 ng/ml BMP4(p≤0.01), as well as between 5 ng/ml and 0, 0.01, 0.1, 25, 50 and 100 ng/ml BMP4(p≤ 0.02). Also, significant differences were observed in proliferation rates between 5ng/ml and 0, 0.01, 0.1, 1, 25 and 100 ng/ml BMP4 (p≤0.01), 25 ng/ml and 0.01, 1 and5 ng/ml BMP4 (p≤0.01), as well as between 50 ng/ml and 0.01 and 0.1 ng/ml BMP4(p≤0.001).Conclusion: The results suggest that addition of 5ng/ml BMP4 had the best effects onthe proliferation and viability rates of CCE mouse ESCs

The changes in morphology and morphometrical indices of endometrium of ovariectomized mice in response to exogenous ovarian hormones

Background: The preparation of endometrium for embryo reception and implantation are controlled by ovarian hormones. These hormones have distinct cyclical changes during estrus cycle. Objective: The aim of this study was to evaluate the changes in morphology and morphometrical indices of endometrium by daily injections of estrogen and progesterone in ovariectomized mouse. Materials and Methods: In total 60 adult NMRI female mice were ovariectomized and after two weeks, they were randomly divided into five groups: control, sham group, estrogen treated mice (which received daily dosage of 0.5 ml/mouse of hormone for five days), progesterone treated mice (which received daily dosage of 0.2 ml/mouse of progesterone hormone for five days) and estrogen-progesterone treated mice (they received 0.5 ml/mouse estrogen on the first day and 0.2 ml/mouse progesterone injections from the second day to the fifth day of treatment). The mice were sacrificed in every day (n=5) up to five days after treatment and their uterine horns were obtained and processed for morphological and morphometrical studies. Results: On the second day of treatment, the diameter of glands was observed to be more in the progesterone group (53.75±6.32μ) than this in the estrogen (45.13±7.78 μ) and estrogen-progesterone treated groups (48.17±13.58 μ). While, the number of glands (76.25±17.37) and thickness of endometrium (39.58±3.37 μ) were observed to be more in the estrogen treated group (p=0.01). Conclusion: Progesterone had effect on the gland whereas estrogen caused increased in height of surface epithelium of endometrium. Overall, the day 2 after treatment (in all experimental groups) is suitable day for sampling for further studies