Archive for the ‘Follicle-Stimulating Hormone’ Category


The effects of insulin plus FSH could have been mediated by either insulin or insulin-like growth factor-1 (IGF-1) receptors or both. However, growing oocytes in FSH plus IGF-1 (10 ng/ml) did not have either deleterious or beneficial effects on oocyte developmental competence (data not shown). Moreover, culture of complexes with FSH and both IGF-1 and insulin together had the same deleterious effect on oocyte developmental competence as culture with FSH plus insulin (data not shown). Thus the deleterious effects of insulin, in combination with FSH, may be mediated via insulin receptors and not by IGF-1 receptors. Buy Advair Diskus Online

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  • RESULTS(3)

    To investigate the relationship between FSH and insulin with respect to the acquisition of oocyte developmental competence in more detail, oocyte-granulosa cell complexes from preantral follicles were cultured for 10 days in control medium (no FSH or insulin), FSH (5 ng/ml) without insulin, insulin (5 ^g/ml) without FSH, or 5 ng/ml FSH plus 0.05-5 ^g/ml insulin. There was no difference in the acquisition of competence to undergo either fertilization and cleavage to the 2-cell stage or the transition from 2cell stage to blastocyst when complexes were cultured in medium without either FSH or insulin, or with FSH alone (Fig. 4). ampicillin antibiotic

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  • While FSH treatment of developing oocyte-granulosa cell complexes significantly reduced the percentage of oocytes competent to undergo fertilization and cleavage to the 2-cell stage (Fig. 2A), the most profound, and unexpected, effects were seen in the oocytes’ competence to develop from the 2-cell stage to blastocyst stage (Fig. 2B). When oocyte-granulosa cell complexes were cultured in control medium (no FSH), 63% of the 2-cell stage derived from control oocytes developed to the blastocyst stage. This percentage was reduced to approximately 20% when the complexes were cultured with 0.5-5 ng/ml FSH (Fig. 2B). antibiotics levaquin

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  • RESULTS(1)Effect of FSH on the Developmental Competence of Oocytes Grown In Vitro in Serum-Free Medium

    As described above, the standard serum-free medium used in this laboratory for the culture of oocyte-granulosa cell complexes from preantral follicles is supplemented with ITS, because complexes cultured in serum-free medium without ITS are very fragile and difficult to maintain throughout the 10-day culture period. Therefore, in the first set of experiments, oocyte-granulosa cell complexes were cultured for 10 days in ITS-supplemented medium, with or without FSH, at concentrations ranging from 0.1 to 5 ng/ ml. No consistent effect of FSH treatment was observed on oocyte growth; the final median oocyte diameter, excluding the zona pellucida, was 70-74 ^m in all groups. antibiotic levaquin

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  • Measurement of LH Receptor (LHR) mRNA Steady State Expression

    The steady-state expression of LHR mRNA in approximately 200 cultured complexes was determined by RNase protection assay exactly as described previously. Protected RNA-RNA hybrids were analyzed by electrophoresis using 6% urea-polyacrylamide gels that were dried and exposed to Fuji phosphor imaging plates and quantified using the Fuji imaging system (Fuji Medical Systems USA, Stamford, CT). The background that was subtracted was the value of an area just below the protected target band and equal in area. The steady-state level of LHR mRNA expression was normalized to the expression of ribosomal protein L19 (Rpl-19) mRNA. All groups within an experiment were cultured and assessed by RNase protection assay at the same time.

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  • MATERIALS AND METHODS(4)

    Evaluation of Expansion (Mucification) of Oocyte-Associated Granulosa Cells

    Oocyte-granulosa cell complexes were removed from the membranes after 10 days of culture and incubated in MEM containing either 100 ng/ml FSH, 1 ^g/ml LH, or control medium for 15 h. Since a component of serum is required for expansion, media were supplemented with 5% FBS (HyClone, Logan, UT). Expansion was scored subjectively as described in detail previously. Intermediate stages of expansion were rarely seen, so complexes were scored as either expanded or not. Data are presented as the percentage of expanded complexes in three independent experiments with at least 50 complexes scored in each experiment. birth control yasmin

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  • After the incubation for oocyte maturation, the granulosa cells were removed from oocytes by drawing the complexes in and out of a Pasteur pipette. The granulosa cell-free oocytes that had undergone germinal vesicle breakdown (GVB), indicative of the resumption of meiosis, were collected and washed three times in fertilization medium. The oocytes that had not undergone GVB were cultured for an additional 24 h in Minimum Essential Medium (MEM) without FSH to assess whether further culture without granulosa cells would allow additional oocytes to undergo GVB. The number of oocytes that underwent GVB with granulosa cells plus the number that underwent GVB subsequently without granulosa cells equaled the total number of oocytes that had developed competence to undergo GVB during the 10-day culture period. Ova were fertilized and preimplantation embryos were cultured as described previously. buy flovent inhaler

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  • MATERIALS AND METHODS(2)

    The culture medium was Waymouth medium MB752/1 supplemented with 0.23 mM pyruvic acid, 50 mg/L streptomycin sulfate, 75 mg/L penicillin G (Sigma), 3 mg/ml BSA (crystallized; ICN Biochemicals, Aurora, OH), ITS (Collaborative Research), and 1 mg/ml fetuin. The oocyte develops within a ball of cells atop a stalk of granulosa cells that is attached to the membrane. Between 200 and 300 complexes, derived from 2 mice, were grown on each membrane, and care was taken to establish the cultures without contact between the complexes. Approximately 90% of the complexes in all groups survived through the 10-day culture period. Cultures were incubated at 37°C in modular incubation chambers (Billups Rothenberg, Del Mar, CA) thoroughly infused with a gas mixture composed of 5% O2:5% cO2:90% N2. Cultures were fed every 2 days by replacement of approximately half of the medium in the compartment below the membrane. All control and experimental groups were isolated and cultured simultaneously for each experiment. buy yasmin online

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