Effect of Angiotensin on the quality of In Vitro Produced [IVP] sheep embryos and expression of Na[+] /K[+] /ATPase

Background: The presence of rennin-angiotensin components in mammalian ovaries and their involvement in ovarian physiology have been established. In the present study, effects of angiotensin II [Ang II] on sodium-potassium adenosine triphosphatase [Na[+] /K[+] /ATPase] expression and development of sheep embryos was evaluated

Methods: The abattoir-derived Cumulus Oocyte Complexes [COC] were randomly allocated into three experimental groups; group I] in vitro Maturation [IVM] of oocytes in the presence of Ang II followed by in vitro fertilization [IVF]/in vitro Culture [IVC] [IVM group], group II] IVM/IVF of oocytes followed by IVC wherein the embryos were exposed to Ang II on day 4 of IVC [D4 group], and group III] IVM/IVF and IVC of oocytes without any angiotensin [Control]. The blastocyst and hatching rates were recorded on days 6 to 8. Day 8 embryos were immunostained with primary and secondary antibodies against Na[+] /K[+] /ATPase alpha 1 and beta 1 subunits

Results: Addition of Ang II during IVM and IVC significantly increased the hatching rate of blastocysts on day 8 compared to the control. The trophectoderm and total blastocyst cells' numbers were significantly increased by addition of Ang II to the IVM and IVC media, though the expression of Na[+] /K[+] /ATPase alpha 1 and beta 1 subunits were positively influenced by the addition of Ang II on day 4 [D4 group]

Conclusion: In conclusion, it seems Ang II through positive effects on embryos, expressed as the greater hatching rate and blastocyst cell number, could increase the sheep embryo developmental rate. These improvements might be partly related to the greater expression of Na[+] /K[+] /ATPase alpha 1 and beta 1 subunits when Ang II was added during IVC

Models predicting success of infertility treatment: a systematic review

Background: Infertile couples are faced with problems that affect their marital life. Infertility treatment is expensive and time consuming and occasionally isn't simply possible. Prediction models for infertility treatment have been proposed and prediction of treatment success is a new field in infertility treatment. Because prediction of treatment success is a new need for infertile couples, this paper reviewed previous studies for catching a general concept in applicability of the models

Methods: This study was conducted as a systematic review at Avicenna Research Institute in 2015. Six data bases were searched based on WHO definitions and MESH key words. Papers about prediction models in infertility were evaluated

Results: Eighty one papers were eligible for the study. Papers covered years after 1986 and studies were designed retrospectively and prospectively. IVF prediction models have more shares in papers. Most common predictors were age, duration of infertility, ovarian and tubal problems

Conclusion: Prediction model can be clinically applied if the model can be statistically evaluated and has a good validation for treatment success. To achieve better results, the physician and the couples' needs estimation for treatment success rate were based on history, the examination and clinical tests. Models must be checked for theoretical approach and appropriate validation. The privileges for applying the prediction models are the decrease in the cost and time, avoiding painful treatment of patients, assessment of treatment approach for physicians and decision making for health managers. The selection of the approach for designing and using these models is inevitable

evaluation of the historical importance of fertility and its reflection in ancient mythology

Myths are reflective of human concerns and needs during ancient times. By reviewing them, it turns out that many human problems today, have a historical background. Among the main themes of ancient mythologies, fertility and reproduction have various representations in ancient civilizations. The purpose of this paper was to review myths and common symbols of fertility and reproduction in ancient civilizations and evaluate the reasons of their continuous importance in different cultures. The data in this review study was obtained by scrutinizing the related literature. The gathered data indicated the multiplicity and variety of fertility symbols in ancient myths. Most ancient fertility symbols were inspired by the nature and some of them like earth and water were common in mythology of different civilizations. Therefore, the symbols consolidate the concept of conformity between human reproductive concerns and the nature's necessities

effect of media supplementation with angiotensin on developmental competence of ovine embryos derived from vitrified-warmed oocytes

Background: This study was aimed to assess the effects of angiotensin II [Ang II] supplementationto the In Vitro Maturation [IVM] and In Vitro Culture [IVC] media ofvitrified-warmed ovine oocytes on their developmental competence and expression ofNa +/K +/ATPase in resulting embryos

Methods: The slaughterhouse-derived immature oocytes [n=1069] were randomly distributedinto four experimental groups: groups I and II] IVM/IVF and IVC of fresh andvitrified oocytes without angiotensin supplementation [Control-Fresh and Control-Vitgroups, respectively]; group III] IVM of vitrified oocytes in the presence of Ang II followedby IVF/IVC [Vit-IVM group]; and group IV] IVM/IVF of vitrified oocytes followedby IVC wherein the embryos were exposed to Ang II on day 4 of IVC [Vit-D4 group].The embryos were immunostained with primary antibodies against Na +/K +/ATPasealpha 1andbeta 1 subunits

Results: In Vit-IVM and Vit-D4 groups, the rates of expanded and total blastocysts onday 7 as well as the proportion of blastocysts on day 8 were increased. The expressionof Na +/K +/ATPasealpha 1 andbeta 1 subunits were positively influenced by the addition of AngII on day 4 [Vit-D4 group].Conclusion: The addition of Ang II to the IVM and IVC media could improve blastocystsformation in vitrified sheep oocytes. This improvement might be related to thegreater expression of Na +/K +/ATPasealpha 1 andbeta 1 subunits when Ang II was added duringIVC

Exogenous secreted frizzled-related protein-4 modulates steroidogenesis of rat granulosa cells through Wnt/beta-catenin and PI3K/AKT signaling pathways

Background: It has been reported that secreted frizzled-related protein-4 known as an antagonist of Wnt signaling pathway plays a role in luteinization process of rodent granulosa cells. The purpose of this study was twofold: 1] to determine whether recombinant human secreted frizzled-related protein-4 [rhSFRP-4] could directly induce terminal differentiation of rat Granulosa Cells [GCs] and 2] to understand how the modulation of beta-catenin and Protein Kinase B [PKB]/AKT activity by exogenous SFRP-4 could be involved in steroidogenesis

Methods: GCs were firstly stimulated with Follicle-Stimulating Hormone [FSH] named as FSH-primed cells then were treated with luteinizing hormone [LH]. Then estradiol [E[2]] and progesterone [P[4]] production levels were assessed in the absence or presence of rhSFRP-4 treatment. The expression levels of activated beta-catenin, pAKTser[473], pGSK3 beta ser[9] were assessed by western blot or immuno-fluoresence

Results: In the presence of rhSFRP-4, there was 38% decreased E[2] levels compared to untreated FSH-primed cells [p<0.05], and P[4] production subsequently decreased. However, in GCs pre-treated with rhSFRP-4 prior to addition of FSH, P[4] levels increased 2-fold compared with untreated cells [p<0.05]. Unexpectedly, treatment with rhSFRP-4 prior to LH stimulation inhibited LH-induced P[4] secretion. Treatment with low [0.5 ng/ml] but not high [50 ng/ml] concentrations of rhSFRP-4 led to significantly increased levels of pGSK3 beta ser[9] [1.6-fold] and nuclear active beta-catenin [2.8-fold] in GCs compared with untreated cells. Interestingly, pre-treating GCs with rhsFPR4 prior to LH stimulation resulted in a 38% decrease in pAKTser[473] levels compared with those in LH-treated cells [p<0.05]

Conclusion: Taken together, our results showed that rhSFRP-4 could directly induce terminal differentiation in GCs via the modulation of beta-catenin and PKB/AKT pathways and that it does so in a dose-dependent manner

Effect of administration of single dose GnRH agonist in luteal phase on outcome of ICSI-ET cycles in women with previous history of IVF/ICSI failure: a randomized controlled trial

GnRH agonist administration in the luteal phase has been suggested to beneficially affect the outcome of intracytoplasmic sperm injection [ICSI] and embryo transfer [ET] cycles. This blind randomized controlled study evaluates the effect of GnRH [Gonadotropine Releasing Hormone] agonist administration on ICSI outcome in GnRH antagonist ovarian stimulation protocol in women with 2 or more previous IVF/ICSI-ET failures. One hundred IVF failure women who underwent ICSI cycles and stimulated with GnRH antagonist ovarian stimulation protocol, were included in the study. Women were randomly assigned to intervention [received a single dose injection of GnRH agonist [0.1 mg of Decapeptil] subcutaneously 6 days after oocyte retrieval] and control [did not receive GnRH agonist] groups. Implantation and clinical pregnancy rates were the primary outcome measures. Although the age of women, the number of embryos transferred in the current cycle and the quality of the transferred embryos were similar in the two groups, there was a significantly higher rate of implantation [Mann Whitney test, p=0.041] and pregnancy [32.6% vs. 12.5%, p=0.030, OR=3.3, 95%CI, 1.08 to 10.4] in the in-tervention group. Our results suggested that, in addition to routine luteal phase support using progesterone, administration of 0.1 mg of Decapeptil 6 days after oocyte re-trieval in women with previous history of 2 or more IVF/ICSI failures led to a signif-icant improvement in implantation and pregnancy rates after ICSI following ovarian stimulation with GnRH antagonist protocol

Embryo donation in Iranian legal system: a critical review

Recent developments and newly-discovered methods for infertility treatment including in vitro fertilization and third party reproduction raise many questions and challenges with different ethical, legal, sociological and psychological dimensions. In Islamic countries, despite great developments in using this technology, the questions concerned with recognition of IVF methods and third party reproduction and legalization of this method are still the challenging ones. The approach of a few Shiite clerics to this issue has facilitated the legalisation of infertility treatment in Iran. The Iranian Parliament, with reference to Shiite clerics' opinions [Fatwa], enacted the Act concerning Embryo Donation to Infertile Couples and its bylaw which can be considered as a successful example of legalization of third party reproduction in an Islamic country. The aforementioned Act permits embryo donation through artificial insemination from legally married couples to infertile couples. However, many of the legal aspects of this event are not specified in this Act and in many cases it added several uncertainties. This uncertainty, especially regarding the rights and duties of recipients and the child, causes important problems which generate more concerns. This article aimed to review the advantages and flaws of the Act. It is believed that the enactment of the aforementioned Act is an important step but an insufficient measure in this field. Important issues have been left unanswered and unclear in this Act which should be considered by legislators in any future revision of it

ART outcomes in GnRH antagonist protocol [flexible] and long GnRH agonist protocol during early follicular phase in patients with polycystic ovary syndrome: a randomized clinical trial

Since increased LH in the early follicular phase in PCOS patients especially in GnRH antagonist protocol could be associated with reduced oocyte quality and pregnancy and impared implantation. The current study was conducted to determine ART outcomes in GnRH antagonist protocol [flexible] and long GnRH agonist protocol and compare them with adding GnRH antagonist in GnRH antagonist [flexible] protocol during early follicular phase in patients with polycystic ovary syndrome undergoing ICSI. In this randomized clinical trial, 150 patients with polycystic ovary syndrome undergoing ICSI were enrolled from 2012 to 2014 and randomly assigned to receive either GnRH antagonist protocol during early and late follicular phase or GnRH antagonist protocol [flexible] or long GnRH agonist protocol. The clinical and laboratory pregnancy in three groups was determined and compared. In this context, the chi-square and Fisher's exact test and ANOVA were used for data analysis. Statistical significance was defined as p<0.05. There was no statistically significant difference with respect to chemical pregnancy and clinical pregnancy between the three groups. Also, other indices such as number and quality of oocytes and embryos were alike. Totally, according to our results, GnRH antagonist protocol during early and late follicular phase and GnRH antagonist protocol [flexible] and long GnRH agonist protocol in patients with polycystic ovary syndrome undergoing ICSI are similarly effective and use of each one based on patients' condition and physicians' opinion could be considered

Experiences of infertile women seeking assisted pregnancy in Iran: a qualitative study

Assisted reproductive technologies [ARTs] are complicated and stressful techniques and the social and cultural norms are major obstacles against their use. Many qualitative studies have been done in the field of women's experiences of infertility, but less is known about the experiences of infertile women seeking assisted pregnancy. The aim of this study was to understand and describe the experience of women who have used assisted reproductive technologies for their current pregnancy. This qualitative study was conducted based on a content analysis approach. With purposive sampling, 12 pregnant women who were using ART were recruited from Avicenna Fertility Center in Tehran. Women were selected purposefully and with maximum variation. Interviews were performed after a positive test of pregnancy and women were introduced to researchers in their first visit of pregnancy in the prenatal clinic. Interviews were recorded, transcribed verbatim and analyzed concurrently. Semi-structured interviews were coded, categorized and the themes were also identified. Four main themes were uncovered which included struggle to achieve pregnancy, fear and uncertainty, escape from stigma and the pursuit to achieve husband satisfaction. It is essential for these women to be counseled and prepared by their health care providers after the use of ARTs. Distress can be reduced for infertile women seeking assisted pregnancy when they are prepared for possible failures, empowered to deal with stigma, and have their partners' involvement in counseling sessions

effect of Setarud [IMOD[TM]] on angiogenesis in transplanted human ovarian tissue to nude mice

One of the promising methods in fertility preservation among women with cancer is cryopreservation of ovarian cortex but there are many drawbacks such as apoptosis and considerable reduction of follicular density in the transplanted ovary. One solution to reduce ischemic damage is enhancing angiogenesis after transplantation of ovarian cortex tissue. The aim of this study was to investigate the effect of Setarud, on angiogenesis in transplanted human ovarian tissue. In this case control study, twenty four nude mice were implanted subcutaneously, with human ovarian tissues, from four women. The mice were randomly divided into two groups [n=12]: the experimental group was treated with Setarud, while control group received only vehicle. Each group was divided into three subgroups [n=4] based on the graft recovery days post transplantation [PT]. The transplanted fragments were removed on days 2, 7, and 30 PT and the expression of Angiopoietin-1, Angiopoietin-2, and Vascular endothelial growth factor at both gene and protein levels and vascular density were studied in the grafted ovarian tissues. On the 2[nd] and 7[th] day PT, the level of Angiopoietin-1 gene expression in case group was significantly lower than that in control group, while the opposite results were obtained for Angiopoietin-2 and Vascular endothelial growth factor. These results were also confirmed at the protein level. The density of vessels in Setarud group elevated significantly on day 7 PT compared to pre-treatment state. Our results showed that administration of Setarud may stimulates angiogenesis in transplanted human ovarian tissues, although further researches are needed before a clear judgment is made

Attitudes about sex selection and sex preference in Iranian couples referred for sex selection technology

Gender preference is prevalent in some communities and using medical techniques to choose the baby's sex may cause the gender discrimination and gender imbalance in communities. Therefore, evaluating the gender preferences and attitudes towards using sex selection technologies seems to be necessary. This cross-sectional study was conducted in Avicenna Fertility Center. Participants were 100 women with one child who were referred for sex selection. Data were collected through self-developed questionnaires. The questions were designed by the researchers at the experts' panel. To determine the validity of the questionnaire, the viewpoints of professors specialized in these issues were obtained. The statistical analysis of the data was performed using SPSS software [Version 11.5], and p<0.05 was considered significant. Tendency toward the male was more than female sex [55.5% male, 15.5% female and 28.5% no tendency]. Majority of participants agreed with sex selection with medical reason and sex selection in order to balance the family. Women's level of education had positive effect on agreements to fetal sex selection with medical and non-medical reasons [p<0.001]. Although gender preferences were toward the male sex but this preference was not very strong. Most participants agreed with non-medical sex selection for balancing the sex composition of their children. It doesn't seem that non-medical sex selection for family balancing causes severe sex imbalance in Iran

effect of sortilin silencing on ovarian carcinoma cells

Our preliminary data on the protein expression of SORT1 in ovarian carcinoma tissues showed that sortilin was overexpressed in ovarian carcinoma patients and cell lines, while non-malignant ovaries expressed comparably lower amount of this protein. In spite of diverse ligands and also different putative functions of sortilin [NTR3], the function of overexpressed sortilin in ovarian carcinoma cells is an intriguing subject of inquiry. The aim of this study was, therefore, to investigate the functional role of sortilin in survival of ovarian carcinoma cell line. Expression of sortilin was knocked down using RNAi technology in the ovarian carcinoma cell line, Caov-4. Silencing of SORT1 expression was assessed using real-time qPCR and Western blot analyses. Apoptosis induction was evaluated using flow cytometry by considering annexin-V FITC binding. [3H]-thymidine incorporation assay was also used to evaluate cell proliferation capacity. Real-time qPCR and Western blot analyses showed that expression of sortilin was reduced by nearly 70-80% in the siRNA transfected cells. Knocking down of sortilin expression resulted in increased apoptosis [27.5 +/- 0.48%] in siRNA-treated ovarian carcinoma cell line. Sortilin silencing led to significant inhibition of proliferation [40.1%] in siRNA-transfected Caov-4 cells as compared to mock control-transfected counterpart [p<0.05]. As it was suspected from overexpression of sortilin in ovarian tumor cells, a cell survival role for sortilin can be deduced from these results. In conclusion, the potency of apoptosis induction via silencing of sortilin expression in tumor cells may introduce sortilin as a potential candidate for developing a novel targeted therapy in patients with ovarian carcinoma

High omega-3: omega-6 fatty acids ratio increases fatty acid binding protein 4 and extracellular secretory phospholipase A2IIa in human ectopic endometrial cells

Endometriosis, a common chronic inflammatory disorder, is defined by the atypical growth of endometrium- like tissue outside of the uterus. Secretory phospholipase A2 group Ha [sPLA2-IIa] and fatty acid binding protein4 [FABP4] play several important roles in the inflammatory diseases, Due to reported potential anti-inflammatory effects of omega-3 and omega-6 fatty acids, the purpose of the present study was to investigate the effects of omega-3 and omega-6 polyunsaturated fatty acids [PUFAs] on fatty acid binding protein 4 and extracellular secretory phospholipase A2IIa in cultured endometrial cells. Ectopic and eutopic endometrial tissues obtained from 15 women were snap frozen. After thawing and tissue digestion, primary mixed stromal f and endometrial epithelial cell culture was performed for 8 days in culture mediums supplemented with normal and high ratios of omega-3 and omega-6 PUFA. sPLA2-IIa in the J culture medium and FABP4 level was determined using enzyme immuno assay [EIA] technique. Within ectopic endometrial cells group, the level of cellular FABP4 and extracellular sPLA2-IIa were remarkably increased under high omega-3 PUFA exposure compared with control condition [p=0.014 and p=0.04 respectively]. Omega-3 PUFAs may increase the level of cellular FABP4 and extracellular sPLA2-IIa in ectopic endometrial cells, since sPLAIIa and FABP4 may affect endometriosis via several mechanisms, more relevant studies are encouraged to know the potential effect of increased cellular FABP4 and extracellular sPLA2-IIa on endometriosis

Enrichment of undifferentiated type a spermatogonia from goat testis using discontinuous percoll density gradient and differential plating

The well documented source for adult multipotent stem cells is Spermatogonial Stem Cells [SSCs]. They are the foundation of spermatogenesis in the testis throughout adult life by balancing self-renewal and differentiation. The aim of this study was to assess the effect of percoll density gradient and differential plating on enrichment of undifferentiated type A spermatogonia in dissociated cellular suspension of goat testes. Additionally, we evaluated the separated fractions of the gradients in percoll and samples in differential plating at different times for cell number, viability and purification rate of goat SSCs in culture. Testicular cells were successfully isolated from one month old goat testis using two-step enzymatic digestion and followed by two purification protocols, differential plating with different times of culture [3, 4, 5, and 6 hr] and discontinuous percoll density with different gradients [20, 28, 30, and 32%]. The difference of percentage of undifferentiated SSCs [PGP9.5 positive] in each method was compared using ANOVA and comparison between the highest percentage of corresponding value between two methods was carried out by t-test using Sigma Stat [ver. 3.5]. The highest PGP9.5 [94.6 +/- 0.4] and the lowest c-Kit positive [25.1 +/- 0.7] in Percoll method was significantly [p

Molecular and serologic diagnostic approaches; the prevalence of herpes simplex in idiopathic men infertile

Human pathogens that can cause infertility may also affect sperm count and quality. Viral infections can be considered as direct and/or indirect cause of male factor infertility. Our goal was to investigate the prevalence of herpes simplex virus in the semen of infertile men attending the Avicenna Infertility Clinic, and to compare it with the herpes virus serology results. This cross sectional study was conducted during 2009-2010. Infertile men participating without any clinical signs of infection with herpes simplex virus, and no obvious cause for their infertility were included. Semen and blood samples were used for Polymerase Chain Reaction [PCR] and serologic testing for these people. Two samples were collected: one ml semen sample to verify the existence of genital herpes simplex virus in infertile men, and blood samples of 217 individuals tested for antibodies to herpes simplex virus. Data were analyzed by SPSS 16. According to the PCR results of semen samples the prevalence of herpes simplex in semen was 12% and serologic test showed 3.2% prevalence within blood. Nine to 10% of IgM negative were PCR positive and only 2-3% of IgM positive were PCR positive. Between herpes serologic studies with positive controls and negative controls by using both tests, there was a significant positive relationship [r=0.718 and p<0.001]. The relationship between semen PCR test results and serological survey of herpes patients with a negative control in both Pearson and Spearman tests was positive and significant [r=0.229 and p=0.001]. Correlation between the PCR results of semen samples with two positive control subjects and a positive IgM test was statistically confirmed [r=0.235 and p<0.001]. We recommend that if there is suspicion to herpes simplex as a microorganism that theoretically could impact semen parameters and cause infertility it is prudent to use PCR technique on semen sample rather than ELISA on serum

Propagation of human germ stem cells in long-term culture

Spermatogonial stem cells [SSCs], a subset of undifferentiated type A spermatogonia, are the foundation of complex process of spermatogenesis and could be propagated in vitro culture conditions for long time for germ cell transplantation and fertility preservation. The aim of this study was in vitro propagation of human spermatogonial stem cells [SSCs] and improvement of presence of human Germ Stem Cells [hGSCs] were assessed by specific markers POU domain, class 5, transcription factor 1 [POU5F1], also known as Octamer-binding transcription factor 4 [Oct-4] and PLZF [Promyelocytic leukaemia zinc finger protein]. Human testicular cells were isolated by enzymatic digestion [Collagenase IV and Trypsin]. Germ cells were cultured in Stem-Pro 34 media supplemented by growth factors such as glial cell line-derived neurotrophic factor, basic fibroblast growth factor, epidermal growth factor and leukemia inhibitory factor to support self-renewal divisions. Germline stem cell clusters were passaged and expanded every week. Immunofluorecent study was accomplished by Anti-Oct4 antibody through the culture. The spermatogonial stem cells genes expression, PLZF, was studied in testis tissue and germ stem cells entire the culture. hGSCs clusters from a brain dead patient developed in testicular cell culture and then cultured and propagated up to 6 weeks. During the culture Oct4 were a specific marker for identification of hGSCs in testis tissue. Expression of PLZF was applied on RNA level in germ stem cells. hGSCs indicated by SSCs specific marker can be cultured and propagated for long-term in vitro conditions

Prevalence of intratubular germ cell neoplasia in cryptorchid testes of infertile men

Cryptorchidism is a common malformation in neonates; surgery or medical treatments are applied during childhood. Untreated cryptorchid testes are in the risk of intratubular germ cell neoplasia [IGCN] and consequently invasive testicular tumors which could be shown by immunohistochemistry staining for placental like acid phosphatase [PLAP] marker. We designed this study to know the prevalence of IGCN in untreated cryptorchid testes of infertile men, in our infertility center as a refferal center. In this cross-sectional study we assessed H and E slides of testicular samples of 13 adult infertile patients with impalpable intra-abdominal testes seeking infertility treatment; then we stained them by PLAP marker. Three [23.08%] samples were positive for PLAP marker means presence of IGCN in testis. One of them showed seminoma besides IGCN. According to the results of this study and the fact that there are adult untreated cryptorchid patients in our country yet, it is suggested to pay more attention in clinical examination, assessment and follow up of such patients for malignancy screening

Evaluation of an aqueous-ethanolic extract from Rosmarinusa officinalis [Rosemary] for its activity on the hormonal and cellular function of testes in adult male rat

Rosmarinus officinalis has been used in traditional medicine extensively. This study evaluated the hormonal and cellular effects of Rosmarinus officinalis extract on testes of adult rats. Thirty male Wistar rats [in three groups] received 50 or 100 mg/Kg b.w of Rosmarinus officinalis extract [made from the plant's leaves, flower and stem] [treatment groups] and 10 mL/Kg b.w normal saline [control group] respectively, on a daily bases by gavage route for 60 days. Then, spermatological properties, histometric parameters and sperm dynamics, testis and body weight, testicular cell population and serum testosterone level were analyzed by an acceptable method. Results showed that the mean serum testosterone level was decreased significantly in both treatment groups [50 and 100 mg/Kg b.w] during the experiment time, compared with control group [p < 0.05]. However, Rosmarinus officinalis did not change the total count, motility and viability of sperm. In addition, Rosmarinus officinalis at both doses did not change body and testes weight and their ratio. Furthermore, Rosmarinus officinalis increased the number of Spermatogonia at both doses, Spermatocyte at doses of 50 mg/Kg b.w, Leydig cell and Spermatid at dose of 100 mg/Kg b.w significantly [p < 0.05]. Rosmarinus officinalis did not significantly affect the number of Spermatozoid and Sertoli cells. In conclusion, it seems that Rosmarinus officinalis may have some hormonal and cellular effects on the testes which can contribute the spermatogenesis process in rat. Rosmarinus officinalis may have antiandrogenic effect potentially indicating the possibility of developing herbal male contraceptive

Comparison of proliferative and multilineage differentiation potential of sheep mesenchymal stem cells derived from bone marrow, liver, and adipose tissue

Despite major progress in our general knowledge related to the application of adult stem cells, finding alternative sources for bone marrow Mesenchymal Stem Cells [MSCs] has remained to be challenged. In this study successful isolation, multilineage differentiation, and proliferation potentials of sheep MSCs derived from bone marrow, adipose tissue, and liver were widely investigated. The primary cell cultures were prepared form tissue samples obtained from sheep 30-35 day fetus. Passage-3 cells were plated either at varying cell densities or different serum concentrations for a week. The Population Doubling Time [PDT], growth curves, and Colony Forming Unit [CFU] of MSCs was determined. The stemness and trilineage differentiation potential of MSCs were analyzed by using molecullar and cytochemical staining approaches. The data was analyzed through one way ANOVA using SigmaStat [ver. 2]. The highest PDT and lowest CFU were observed in adipose tissue group compared with other groups [p<0.001]. Comparing different serum concentrations [5, 10, 15, and 20%], irrespective of cell sources, the highest proliferation rate was achieved in the presence of 20% serum [p<0.001]. Additionally, there was an inverse relation between cell seeding density at culture initiation and proliferation rate, except for L-MSC at 300 cell seeding density. All three sources of fetal sheep MSCs had the identical trilineage differentiation potential. The proliferative capacity of liver and bone marrow derived MSCs were similar at different cell seeding densities except for the higher fold increase in B-MSCs at 2700 cells/cm2 density. Moreover, the adipose tissue derived MSCs had the lowest proliferative indices

vitro culture of human testicular stem cells on feeder-free condition

Spermatogonial stem cells are subpopulation of spermatogonial cells in testis tissue that support beginning and maintenance of spermatogenesis. Ubiquitin carboxy-terminal hydrolase L1 [UCHL1] could be a specific marker for identification of spermatogonial stem cells including spermatogonial sperm cells [SSCs] in testis tissue and during the culture; therefore we undertook this study to culture these human testicular stem cells [hTSCs] in vitro and approved the presence of human testicular stem cells [hTSCs] by UCHL1, also known as PGP9.5. Enzymatic digestion of human testicular biopsies was done by collagenase IV [4 mg/ml] and trypsin [0.25%]. Differential plating of testicular cells in DMEM/F12 and 10% FBS was applied for 16 hr. Floating cells were collected and transferred onto laminin-coated plates with Stem-Pro 34 media supplemented with growth factors of GDNF, bFGF, EGF and LIF to support self-renewal divisions; testicular stem cell clusters were passaged every 14 days for two months. Spermatogonial cells propagation was studied through Expression of UCHL1 in testis tissue and the entire testicular stem cell culture. Testicular stem cell clusters from 10 patients with obstructive azoospermia were cultured on laminin-coated plates and subsequently propagated for two months. The average of harvested viable cells was approximately 89.6%. UCHL1 was expressed as specific marker in testicular stem cells entire the culture. Human testicular stem cells could be obtained from human testicular tissue by a simple digestion, culturing and propagation method for long-term in vitro conditions. Propagation of these cells approved by specific marker UCHL1, during the culture period