correlation between the endometrial integrins and osteopontin expression with pinopodes development in ovariectomized mice in response to exogenous steroids hormones

The ovariectomized animals are good models to evaluate the effect of different steroid hormone treatments on implantation events and the pattern of integrin expression. Therefore, this study was performed to compare the expression of integrins and osteopontin [OPN] in correlation with pinopode development in ovariectomized mice endometrium which was subjected to steroid hormones. Ovariectomized mice were subjected to estrogen, progesterone and estrogen-progesterone hormones. Their uterine horns were evaluated for integrin expression by immunohistochemistry and real-time RT-PCR and for pinopode development by transmission and scanning electron microscopic studies. No immunostaining for integrin and OPN molecules were detected in the endometrium of non-ovariectomized mice except in metestrus phase. The alpha4 and beta 1 integrin genes were expressed in all phases of estrous cycle. In ovariectomized mice, no reaction was detected in the endometrium of control, sham and estrogen-treated groups, but in both progesterone-treated groups, all examined genes were expressed. There was not any correlation between pinopodes and integrin expression in ovariectomized mice. The progesterone is more effective on endometrial integrin expression than estrogen and differences in the expression pattern of integrins reflect their important and different roles in embryo implantation. The pinopodes may have minor effect in mice implantation or have some delay in their expressions in ovariectomized mice which were subjected to exogenous hormones

Effects of different doses of bone morphogenetic protein 4 on viability and proliferation rates of mouse embryonic stem cells

In this study, we examined the effect of different doses of bone morphogenetic protein 4 [BMP4] on CCE mouse embryonic stem cells [ESCs] viability and proliferation rates in order to improve the outcome of induction processes and make a system with highest viability and proliferation rates for further studies on BMP4 roles in multiple developmental stages. Expression of Oct-4 was studied and confirmed in this cell line immunocytochemically. Also, in order to evaluate the proliferation and viability rates 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 and survival rates respectively. Data analysis was done with ANOVA test. The results showed that there were significant differences between the mean percent of viability between 1ng/ml and 0 ng/ml [control] and 50 and 100 ng/ml BMP4 [p

Transplantation and homing of mouse embryonic stem cells treated with erythropoietin in spleen and liver of irradiated mice

The present study was designed to evaluate the homing potential of mouse embryonic stem cells [ESC] treated with erythropoietin [EPO] in hematopoietic organs such as spleen and liver after transplantation using morphological and immuno-histochemical techniques. Day-four embryoid body [EB]-derived cells were dissociated and re-plated in medium in the presence and absence of EPO for three days. The EPO- and untreated differentiated cells were labeled with 5-bromo-2 deoxyuridine [BrdU] before transplantation and analyzed using flow cytometry and reverse transcription-PCR methods. BrdU-labeled cells were injected via the tail vein into irradiated adult mice in both groups. The spleen colony-forming unit assay [CFU-S] was performed 12 days after transplantation. Immuno-histochemistry was also carried out to trace transplanted cells. The percentage of CD34 positive cells was 5.51 +/- 1.06% in the EPO-treated group and 1.63 +/- 0.225% in untreated group. The RT-PCR analysis showed that the EPO-treated cells expressed epsilon globin, beta H1 globin, RUNX1 and EPO receptor genes, but the beta-major globin gene was not expressed. The number of colonies formed in the spleens of treated group [17.33 +/- 4.726] was significantly different from the control group [6 +/- 1]. The population of BrdU positive cells in spleen of EPO-treated cell-transplanted group was higher than that of the control group. Also, BrdU positive cells were observed in the central vein of the liver sections of EPO-treated and control groups but were not observed in the liver parenchyma. There were not BrdU positive cells in the spleen and liver sections of the sham group. Our results confirm that ESC have the ability to home and form colonies in spleen after transplantation and EPO-treated EB-derived cells caused an increase in the number of colonies in spleen after CFU-S