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Showing 12 results for Salehnia

Ali A Movassagh-Pour, Mojdeh Salehnia, Ali A Pourfatollah, Sayed M Moazzeni,
Volume 1, Issue 1 (1-2003)

Background: Embryonic stem cells (ESc) are pluripotent cells which have been used as a model to study the mechanism that control the embryogenesis and early mammalian development in vitro. The aim of this study was to isolate and produce embryonic stem cells from late blastocyst stage embryos in mice. Materials and Methods: Blastocyst stage embryos from pregnant NMRI mice were obtained and cultured for 24 h in DMEM medium. 4-6 days after hatching, the inner cell masses (ICM) formed colonies which were then collected mechanically and trypsinized. Several subcultures were prepared in the medium supplemented with 0.1 mM 2 Mercaptoethanol, 1000 U/ml Leukemia Inhibitory Factor (LIF) and 10% Fetal Bovine Serum (FBS). The (ESc) were recognized by alkaline phosphates histochemistry using azo-coupling method. Results: The results demonstrated that a highly pluripotent stem cell line was derived from the blastocyst stage embryos of NMRI mice; however, the rate of colonies was as low as 10%. Conclusion: The LIF is effective to culture and maintain the isolated ICM colonies in undifferentiated condition in the absence of feeder layer.
Mojdeh Salehnia,
Volume 1, Issue 1 (1-2003)

Background: The aim of this study was to determine the correlation between ultrastructural studies for pinopodes expression after ovarian hyperstimulation and progesterone injection in mice. Materials and Methods: Adult NMRI mice were superovulated using human menopasual gonadotropic (hMG) and human chorionic gonadotropic (hCG) hormones; after that, daily injection of progesterone (1 mg/mouse) was performed. Animals were sacrificed by cervical dislocation 3.5 and 4.5 days after hCG injection. Tissues of uterine horns were obtained and processed for scanning (SEM) and transmission (TEM) electron microscopy studies. The pseudopregnant control samples were studied same as experimental groups. Results: The SEM and TEM observations showed that in control groups on 3.5 days of pregnancy, there were some pinopodes. All apical cell surfaces expressed these projections on the forth day. In progestrone-injected group, well developed pinopods were expressed 3.5 days after hCG injection and they were transformed to small projections on the fourth day following hCG injection. Also, the life span of pinopods was limited to a short time. At the TEM levels, the pinopods were seen as swelling process on the apical surface, which were more pronounced on day 3.5 of hCG injection in hyperstimulated and progestrone injection. Conclusion: The progestrone may cause premature expression of pinopodes and the implantation failure after ovarian induction may be due to these timing changes.
Behnaz Sheikholslami, Mojdeh Salehnia, Mojtaba Rezazadeh,
Volume 2, Issue 1 (7-2004)

The cytokine of granulocyte macrophage colony stimulating factor (GM-CSF) is a glycoprotein, which is synthesized in the female reproductive tract and has embryonic trophic effect in mammals. The objective of this study was to examine the optimal dosage of GM-CSF to improve the mouse embryo development in vitro. To collect two and eight cells embryos, the pregnant NMRI mice were sacrificed by cervical dislocation at 48 h and 72 h post hCG injections, respectively. The embryos were cultured randomlly in T6 medium supplemented with 5 mg /ml bovine serum albumin (BSA) and 0, 2, and 10 ng / ml human rGM-CSF. The data of blastocyst formation and hatching in different groups of embryo culture were compared by chi-square analysis. The results showed that the developmental rates of 2 and 8 cells embryos to hatching blastocyst in the presence of 2 ng/ml of GM-CSF their control groups (51.5% and 49.7%, respectively) were more than those in the other groups, but insignificant. It seems more researches are necessary to confirm this suggestion that the GM-CSF with 2 ng/ml concentration may have a better potential, not only to enhance the developmental rates of 2 and 8 cells embryos but also for decreasing the degeneration of those embryos.
Reza Mahmoudi, Aligholi Subhani, Parichehr Pasbakhsh, Farideh Etesam, Iraj Amiri, Mozhdeh Salehnia, Farid Abolhasani,
Volume 3, Issue 2 (7-2005)

Background: In vitro maturation (IVM) of oocytes is a promising technique to reduce the costs and avert the side-effects of gonadotropin stimulation for in vitro fertilization (IVF). The pregnancy rates from oocytes matured in vitro are much lower than those of in vivo stimulation cycles, indicating that optimization of IVM remains a challenge. Objective: In this study, we investigated the effect of cumulus cells on maturation and fertilization rate of immature oocytes (Germinal vesicle). Materials and Methods: Germinal vesicle (GV) oocytes were recovered from 6-8 weeks old Balb C female mice 48hr after injection of 10 IU pregnant mare serum gonadotropin (PMSG). Collected oocytes were divided into two groups. Group A: GV oocytes without cumulus (denuded oocyte). Group B: GV oocytes with cumulus cells (cumulus-oocyte complex). The oocytes in both groups were cultured in TCM-199 medium in a humidified atmosphere of 5% CO2 in air at 37�C. The maturation, fertilization and developmental rates were recorded after 24hr. Results: Maturation, fertilization and developmental rates in denuded oocytes (DO) were 65.1%, 68.02%, 78.63% respectively, and in cumulus-oocyte complex (COC) were 78.20%, 85.57% and 85.05%, respectively. The maturation, fertilization and developmental rates of COC were significantly higher than those of DO (p<0.05). Conclusion: The results show that cumulus cells have beneficial effects on maturation, fertilization and cleavage rates of mice oocytes.
Mojdeh Salehnia, Mitra Arianmanesh, Mandana Beigi,
Volume 4, Issue 1 (7-2006)

Background: The preparation of endometrium for embryo reception is dependent on the ovarian hormones, which are affected by ovarian hyperstimulation procedure.
Objective: The aim of this study was to evaluate the changes in morphometrical indices of endometrium by the daily injection of progesterone after mouse ovarian induction.
Materials and Methods: Adult virgin female mice were selected and divided into control and experimental groups. Experimental groups were superovulated using human menopasual gonadotropic hormone (HMG), and human chorionic gonadotropic hormone (HCG), then they, were subdivided into two groups, which one group was also injected daily by progesterone. All control and hyperstimulated groups were rendered pseudopregnant by cervical stimulation. Three and four days after the HCG injection, the samples of uterine horns were aparted and processed for light microscopic studies.
Results: Our results showed that in the progesterone-injected group, the height of surface and glandular epithelium was decreased on day three (17.6±3.55, 10.02±2.6) and day four (16.9±4.24, 1.6±0.84) respectively, and it had low columnar morphology in comparison with the hyperstimulated and control groups. Also the intercellular spaces of stroma in progesterone-injected group were narrower than these in the other groups.
Conclusion: Ovarian hyperstimulation followed by progesterone injection alter the morphometrical indices of surface and glandular epithelium of endometrium, which could affect on its receptivity.
Fatemeh Peyghambari, Mojdeh Salehnia, Mehdi Forouzandeh Moghadam, Mojtaba Rezazadeh Valujerdi, Ebrahim Hajizadeh,
Volume 6, Issue 4 (7-2008)

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.
Kamran Haidari, Mojdeh Salehnia, Mojtaba Rezazadeh Valojerdi, Shahram Pourbyranvand,
Volume 6, Issue 5 (7-2008)

Background: The ultrastructural analysis of cultured follicles could direct us to understand subcellular changes during in vitro culture.
Objective: This study was done to verify the ultrastructural characteristics of in vitro cultured mouse isolated preantral follicles in co-culture system in the presence and absence of leukemia inhibitory factor (LIF)
Materials and Methods: Mechanically isolated preantral follicles were divided into four groups: control without LIF, control with LIF, co-cultured group with LIF, co-cultured group without LIF. In co-culture groups the follicles were cultured with cumulus cells. After 4 days the follicles were processed and sectioned for transmission electron microscopic examination.
Results: The oocytes of cultured preantral follicles in all studied groups demonstrated a homogeneous cytoplasm and they had the round or ovoid shaped mitochondria with light matrix and cristae. Their endoplasmic reticulum cisternae were in association with mitochondria and Golgi complex. The cortical granules and the aggregation of mitochondria around the germinal vesicle were prominent in both co-cultured groups. The organelle distribution in granulosa cells was normal in all groups of study and no sign of cell death was observed. In both co-cultured systems the granulosa cells contained mitochondria with tubular cristae, a well developed smooth endoplasmic reticulum and several large lipid droplets, characteristics of steroid synthesis cells. Conclusion: The oocyte and granulosa cells in co-cultured system showed more remarkable maturation features than that of control.
Ali Abedelahi, Mojdeh Salehnia, Allameh Abdolamir, Ebrahim Hajizadeh,
Volume 7, Issue 2 (7-2009)

Background: Many attempts have done to improve cryopreservation of mammalian ovaries using simple, economical and efficient technique “vitrification”.
Objective: The aim of the present study was to compare the mouse ovaries cryopreservation by direct cover vitrification (DCV) using different concentrations of ethylene glycol (EG) with conventional vitrification methods (CV).
Materials and Methods: Ninety NMRI mice were sacrificed by cervical dislocation; their ovaries were divided into three main experimental groups: control or non-vitrified group, CV group and DCV groups with 4, 6 and 8M EG as cryoprotectant. After vitrification-warming, the viability of mechanically isolated follicles and the morphology of ovarian follicles by light and electron microscopes were studied.
Results: The normality of primary and preantral follicles in non-vitrified and CV groups were higher than those achieved by DCV groups (p<0.001). The survival rates of isolated follicles in non-vitrified, CV and DCV groups with 4M, 6M and 8M ethylene glycol were 98.32, 96.26, 84.10, 85.46 and 84.56 %, respectively and in DCV groups it was lower than other groups (p<0.001). The ultrastructure of ovarian follicles was well preserved in CV technique. The follicles in DCV groups appeared to have vacuolated oocyte with nuclear shrinkage and irregular distribution of cytoplasmic organelles. Their mitochondria were located mainly in the sub cortical part of the oocyte and the granulosa cells demonstrated some signs of degeneration.
Conclusion: DCV of mouse ovarian tissue using only EG has induced some alteration on the fine structure of follicles. The integrity of mouse ovarian tissue was affected by DCV technique more than CV.
Sahar Hatami, Saeed Zavareh, Mojdeh Salehnia, Taghi Lashkarbolouki, Mohammad Taghi Ghorbanian, Isaac Karimi,
Volume 12, Issue 1 (2-2014)

Background: Cryopreservation of ovarian tissues and pre-antral follicles is a promising prospect for preservation of women fertility.
Objective: The aim of this study was to evaluate the in vitro developmental competence of mouse vitrified pre-antral follicles in comparison to isolated pre-antral follicles derived from vitrified ovaries in the presence of alpha lipoic acid (ALA).
Materials and Methods: Pre-antral follicles derived from fresh, vitrified-warmed ovarian tissues and vitrified–warmed pre-antral follicles were cultured individually with or without ALA, followed by adding hCG to induce ovulation. The follicle growth, oocyte maturation, and embryo development were assessed.
Results: The diameter and development of follicles, oocyte maturation and embryo development rates were significantly higher in ALA supplemented groups compared to the respective ALA-free conditions groups. Aforementioned parameters were significantly higher in vitrified-warmed follicles in comparison to follicles derived from vitrified-warmed ovaries.
Conclusion: These findings support a superior performance of pre-antral follicles when vitrified rather than when isolated from vitrified ovaries with regard to increasing the rates of developmental parameters. Moreover, ALA improves the in vitro maturation of pre-antral follicles in vitrified and non-vitrified samples. 
Mehri Fayazi, Mojdeh Salehnia, Saeideh Ziaei,
Volume 14, Issue 7 (7-2016)

Background: Stem cell factor (SCF) is a transcriptional factor which plays crucial roles in normal proliferation, differentiation and survival in a range of stem cells. 
Objective: The aim of the present study was to examine the proliferation effect of different concentrations of SCF on expansion of human endometrial CD146+ cells. 
Materials and Methods: In this experimental study, total populations of isolated human endometrial suspensions after fourth passage were isolated by magnetic activated cell sorting (MACS) into CD146+ cells. Human endometrial CD146+ cells were karyotyped and tested for the effect of SCF on proliferation of CD146+ cells, then different concentrations of 0, 12.5, 25, 50 and 100 ng/ml was carried out and mitogens-stimulated endometrial CD146+ cells proliferation was assessed by MTT assay. 
Results: Chromosomal analysis showed a normal metaphase spread and 46XX karyotype. The proliferation rate of endometrial CD146P + P cells in the presence of 0, 12.5, 25, 50 and 100 ng/ml SCF were 0.945±0.094, 0.962±0.151, 0.988±0.028, 1.679±0.012 and 1.129±0.145 respectively. There was a significant increase in stem/ stromal cell proliferation following in vitro treatment by 50 ng/ml than other concentrations of SCF (p=0.01). 
Conclusion: The present study suggests that SCF could have effect on the proliferation and cell survival of human endometrial CD146P+P cells and it has important implications for medical sciences and cell therapies.
Mina Jafarabadi, Mojdeh Salehnia, Rana Sadafi,
Volume 15, Issue 1 (1-2017)

Background: The animal models of endometriosis could be a valuable alternative tool for clarifying the etiology of endometriosis.
Objective: In this study two endometriosis models at the morphological and molecular levels was evaluated and compared.
Materials and Methods: The human endometrial tissues were cut into small fragments then they were randomly considered for transplantation into γ irradiated mice as model A; or they were isolated and cultured up to fourth passages. 2×106 cultured stromal cells were transplanted into γ irradiated mice subcutaneously as model B. twenty days later the ectopic tissues in both models were studied morphologically by Periodic acid-Schiff and hematoxylin and eosin staining. The expression of osteopontin (OPN) and matrix metalloproteinase 2 (MMP2) genes were also assessed using real time RT-PCR. 17-β estradiol levels of mice sera were compared before and after transplantation.
Results: The endometrial like glands and stromal cells were formed in the implanted subcutaneous tissue of both endometriosis models. The gland sections per cubic millimeter, the expression of OPN and MMP2 genes and the level of 17-β estradiol were higher in model B than model A (p=0.03).
Conclusion: Our observation demonstrated that endometrial mesenchymal stromal cells showed more efficiency to establish endometriosis model than human endometrial tissue fragments
Mojdeh Salehnia, Mehri Fayazi, Shokreya Ehsani,
Volume 15, Issue 4 (6-2017)

Background: Concerning the low population of human endometrial mesenchymal cells within the tissue and their potential application in the clinic and tissue engineering, some researches have been focused on their in vitro expansion.
Objective: The aim of this study was to evaluate the effect of leukemia inhibitory factor (LIF) as a proliferative factor on the expansion and proliferation of human endometrial stromal cells.
Materials and Methods: In this experimental study, the isolated and cultured human endometrial stromal cells from women at ovulatory phase aged 20-35 years, after fourth passage were divided into control and LIF-treated groups. In the experimental group, the endometrial cells were treated by 10 ng/ml LIF in culture media and the cultured cells without adding LIF considered as control group. Both groups were evaluated and compared for proliferation rate using MTT assay, for CD90 marker by flow cytometric analysis and for the expression of Oct4, Nanog, PCNA and LIFr genes using real-time RT-PCR.
Results: The proliferation rate of control and LIF-treated groups were 1.17±0.17and 1.61±0.06 respectively and there was a significant increase in endometrialstromal cell proliferation following in vitro treatment by LIF compared to controlgroup (p=0.049). The rate of CD90 positive cells was significantly increased in LIFtreatedgroup (98.96±0.37%) compared to control group (94.26±0.08%) (p=0.0498).Also, the expression ratio of all studied genes was significantly increased in the LIFtreatedgroup compared to control group (p=0.0479).
Conclusion: The present study showed that LIF has a great impact on proliferation, survival, and maintenance of pluripotency of human endometrial stromal cells and it could be applicable in cell therapies.

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