ABSTRACT
Objective: Bone marrow has recently been recognized as a novel source of stem cells for the treatment of wide range of diseases. A number of studies on murine bone mar-row have shown a homogenous population of rare stage-specific embryonic antigen 1 [SSEA-1] positive cells that express markers of pluripotent stem cells. This study focuses on SSEA-1 positive cells isolated from murine bone marrow in an attempt to differentiate them into insulin-secreting cells [ISCs] in order to investigate their differentiation potential for future use in cell therapy
Materials and Methods: This study is an experimental research. Mouse SSEA-1 positive cells were isolated by Magnetic-activated cell sorting [MACS] followed by characterization with flow cytometry. Induced SSEA-1 positive cells were differentiated into ISCs with specific differentiation media. In order to evaluate differentiation quality and analysis, dithizone [DTZ] staining was use, followed by reverse transcription polymerase chain reaction [RT-PCR], immunocytochemistry and insulin secretion assay. Statistical results were analyzed by one-way ANOVA
Results: The results achieved in this study reveal that mouse bone marrow contains a population of SSEA-1 positive cells that expresses pluripotent stem cells markers such as SSEA-1, octamer-binding transcription factor 4 [OCT-4] detected by immunocytochemistry and C-X-C chemokine receptor type 4 [CXCR4] and stem cell antigen-1 [SCA-1] detected by flow cytometric analysis. SSEA-1 positive cells can differentiate into ISCs cell clusters as evidenced by their DTZ positive staining and expression of genes such as Pdx1 [pancreatic transcription factors], Ngn3 [endocrine progenitor marker], Insulin1 and Insulin2 [pancreaticbeta-cell markers]. Additionally, our results demonstrate expression of PDX1 and GLUT2 protein and insulin secretion in response to a glucose challenge in the differentiated cells
Conclusion: Our study clearly demonstrates the potential of SSEA-1 positive cells to differentiate into insulin secreting cells in defined culture conditions for clinical applications
ABSTRACT
Infections are one of the correctable causes of infertility with low cost and cost effective treatment. The 50% of infertile cases is related to men in some way, and 30% of them are absolutely related to them. Mycoplasmas are the smallest microorganisms with capability of DNA replication. Present study is planned to compare the mycoplasma infection in infertile men and men with established fertility. 45 Semen samples were collected from case and control persons who referred to Royan Infertility and Fertility Institute between 2004 and 2005 and stored in -20 degree°C until time of test. DNA was extracted from semen using phenol chloroform. PCR reaction was done by mycoplasma specific primers. Mycoplasma genitalium gene was amplified in 6 [40%] cases from 15 infertile semen samples and 11 [36.6%] from 30 control semen samples. Probability of genital infection, at least, in these studies group, is very lower than other communities' reports
Subject(s)
Humans , Male , Infertility, Male , Fertility , SpermatozoaABSTRACT
The purpose of this study was to evaluated the effect of beta-mercaptoethanol on resumption of meiosis, in vitro maturation of immature mouse oocytes and resulting embryo development with and without BSO [DL-Buthionine sulfoximine] Material and Germinal vesicle [GV] were recovered from 6-8 weeks old NMRI ovaries and cultured in maturation medium in MEMalpha supplemented with 7.5IU/ml hCG, 100mIU/ml rhFSH, 5% FCS [control group] and adding 100 micro m beta-mercaptoethanol [group 1] or with 5mM BSO + 100 micro m beta-mercaptoethanol [group 2] for 24h. The matured oocytes then were fertilized and cultured for 5 days. Fertilization and development were accomplished in T6 medium.The percentage of GV oocytes reaching to metaphase I [or undergo GVBD] were 78.5%, 85%, 86% in control group, group 1 and group 2 respectively, that no significant difference was detected between groups. The proportion of oocytes that progressed to the metaphase II [MII] stage was minimum within 5mM BSO group [group 2] and maximum within beta-mercaptoetanol group [group 1] with significant difference comparing with control and each other [P<=0.05]. The percentage of embryos reaching to morula stage within beta-mercaptoetanol group was significantly higher than the control group [5% and 12.2% respectively]. None of oocytes treated with BSO could pass the 8 cell stage. beta-mercaptoetanol enhances IVM and improves embryo development. While adding BSO into the maturation medium even with beta-mercaptoetanol decreases maturation and declines the embryo development
Subject(s)
Animals , Buthionine Sulfoximine , Mice , Embryonic Development , Oxidative Stress , Apoptosis , OocytesABSTRACT
DEHP [di[2-ethylhexyl] phthalate]] is widely used in plastic industry and some reproductive toxicity has been shown with it. So, this study was designed to evaluate DEHP effects on resumption of meiosis and in vitro maturation of mouse oocytes as well as development of embryos resulted from them. Mice of 4-6 weeks old were administered daily doses of 50, 100, 200 microl of 2.56 micro M DEHP solution for 12 days. Immature mouse oocytes were recovered from all experimental groups and matured in MEM-alpha medium containing 5% FCS with and without 7.5 IU hCG and 100 mIU rFSH. IVF was performed T6 medium. Resumption of meiosis and in vitro maturation were significantly lower in all experimental groups in culture media without hormones compared to controls. Fertilization and embryo development were also significantly decreased in both culture media [with and without hormones]. This study showed the adverse effects of DEHP on in vitro maturation and embryo development in a dose dependent manner