Primary human granulosa cells were obtained with informed patient consent following approval from the University of British Columbia Research Ethics Board. The controlled ovarian stimulation protocol for in vitro fertilization patients consisted of either luteal-phase naferelin acetate (Synarel, Pfizer, Kirkland, Quebec, Canada) or follicular phase GnRH antagonist (Ganirelix; Merck Canada) down-regulation. Gonadotropin stimulation began on menstrual cycle day 2 with human menopausal gonadotropin (hMG; Menopur, Ferring, Canada) and recombinant FSH (Puregon, Merck, Canada) and was followed by human chorionic gonadotropin administration 34–36 hours before oocyte retrieval, based on follicle size. Granulosa cells were purified by density centrifugation from follicular aspirates collected from women undergoing oocyte retrieval as previously described [29 (link), 30 (link)].
Synarel
Manufactured by Pfizer
Sourced in Canada
Synarel is a laboratory equipment product used for the detection and measurement of various analytes. It functions as a specialized device for analytical purposes, providing accurate and reliable data. The core function of Synarel is to facilitate the analysis of samples, enabling researchers and scientists to obtain relevant information for their research and experiments.
Automatically generated - may contain errors
6 protocols using synarel
1
Isolation of Primary Human Granulosa Cells
2
Isolation and Culture of Primary Human Granulosa Cells
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In this study, primary hGL cells were obtained with informed consent from patients following approval from the University of British Columbia Ethics Board. Follicular samples were anonymized promptly post-collection, and each primary culture consisted of cells sourced from a single patient, totaling 20 patients in the study cohort. Two controlled ovarian stimulation protocols were employed for in vitro fertilization patients: (1) luteal-phase nafarelin acetate (Synarel, Pfizer, Kirkland, Quebec, Canada) and (2) follicular phase GnRH antagonist (Ganirelix; Merck Canada) downregulation. Gonadotropin stimulation commenced on menstrual cycle day 2 using human menopausal gonadotropin (hMG; Menopur, Ferring, Canada) and recombinant FSH (Puregon, Merck, Canada). Human chorionic gonadotropin was administrated 34–36 h before oocyte retrieval, based on follicle size. Granulosa cells were purified by density centrifugation from follicular aspirates obtained from women undergoing oocyte retrieval, as previously described [18 (link)]. Each sample of primary hGL cells collected from individual females was cultured separately. The purified hGL cells were seeded at a density of 5 × 105 cells per well in 6-well plates with the same culture environment and medium used for the SVOG cell line.
3
Controlled Ovarian Hyperstimulation for IVF
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Patients undergoing IVF/ET treatment initially received controlled ovarian hyperstimulation using medication that prevents the premature luteinization, including GnRH antagonists (ganirelix, Merck, Canada) in the follicular phase or triptorelin acetate (Synarel, Pfizer, Canada) in the luteal phase of the previous cycle to downregulate the pituitary gonadotropin-releasing hormone receptors and subsequent transduction pathways. On the second day of the menstrual cycle, appropriate human menopausal gonadotropin (hMG, Menopur, Ferring, Canada) or recombinant FSH (Puregon, Merck, Canada) were administered to stimulate follicular growth. When the diameter of the leading follicle reached > 18 mm or the diameters of at least three follicles reached > 17 mm, hCG (Pregnyl, Merck) was administered to trigger final oocyte maturation. Oocytes were retrieved under vaginal ultrasound-guiding 34–36 h after the trigger, and corresponding follicular fluid was collected.
4
Isolation and Culture of Primary Human Granulosa Cells
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Primary hGL cells were obtained with informed patient consent following approval from the University of British Columbia Ethics Board. Two controlled ovarian stimulation protocols for in vitro fertilization patients were utilized: (1) one involved luteal-phase nafarelin acetate (Synarel, Pfizer, Kirkland, Quebec, Canada) and (2) another was based on follicular phase GnRH antagonist (Ganirelix; Merck Canada) down regulation. Gonadotropin stimulation was initiated on menstrual cycle Day 2 by administration of human menopausal gonadotropin (hMG; Menopur, Ferring, Canada) and recombinant FSH (Puregon, Merck, Canada). Based on follicle size, human chorionic gonadotropin was administered 34–36 h before oocyte retrieval. Granulosa cells were purified by density centrifugation of follicular aspirates obtained from women undergoing oocyte retrieval as previously described [49 (link)]. Primary hGL cells collected from individual females were cultured separately. Purified hGL cells were seeded in 6-well plates in the same culture environment and culture medium that was used with the SVOG cell line.
5
Primary Human Granulosa Cell Isolation
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Primary human granulosa cells were obtained with informed patient consent following approval from the University of British Columbia Research Ethics Board. The controlled ovarian stimulation protocol for in vitro fertilization patients consisted of either luteal-phase naferelin acetate (Synarel, Pfizer, Kirkland, Quebec, Canada) or follicular phase GnRH antagonist (Ganirelix; Merck Canada) down-regulation.
Gonadotropin stimulation began on menstrual cycle day 2 with human menopausal gonadotropin (hMG; Menopur, Ferring, Canada) and recombinant FSH (Puregon, Merck, Canada) and was followed by human 5 chorionic gonadotropin administration 34-36 h before oocyte retrieval, based on follicle size. Granulosa cells were purified by density centrifugation from follicular aspirates collected from women undergoing oocyte retrieval, as previously described [28, 29] . Cells were cultured in a humidified atmosphere containing 5% CO 2 at 37 °C with DMEM/F-12 medium supplemented with 10% charcoal/dextran-treated FBS and 1X GlutaMAX for 5 days before treatment [30] .
Gonadotropin stimulation began on menstrual cycle day 2 with human menopausal gonadotropin (hMG; Menopur, Ferring, Canada) and recombinant FSH (Puregon, Merck, Canada) and was followed by human 5 chorionic gonadotropin administration 34-36 h before oocyte retrieval, based on follicle size. Granulosa cells were purified by density centrifugation from follicular aspirates collected from women undergoing oocyte retrieval, as previously described [28, 29] . Cells were cultured in a humidified atmosphere containing 5% CO 2 at 37 °C with DMEM/F-12 medium supplemented with 10% charcoal/dextran-treated FBS and 1X GlutaMAX for 5 days before treatment [30] .
6
Ovarian Stimulation for IVF: Long GnRH Agonist Protocol
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Ovarian stimulation was carried out by the long protocol using gonadotropin-releasing hormone agonist (GnRha), Synarel (Pfizer, Israel) and human menopausal gonadotropin (HMG) (Menopur -Ferring Pharmaceuticals, IL) or recombinant follicle-stimulating hormone (FSH) (Follitropin alpha, Gonal F, Merck Serono IL or follitropin beta, Puregon, MSD IL).
Monitoring was performed by serial measurement of blood 17β-oestradiol and progesterone and vaginal ultrasound scanning of developing follicles. Ovulation triggering was performed with recombinant human chorionic gonadotropin (rHCG) (Ovitrelle 250 mcg, Merck Serono, IL) when two to three leading follicles (18-19 mm in diameter) were demonstrated on ultrasound. Oocyte collection was performed 35-36 h later (Lahav-Baratz et al., 2004) .
Monitoring was performed by serial measurement of blood 17β-oestradiol and progesterone and vaginal ultrasound scanning of developing follicles. Ovulation triggering was performed with recombinant human chorionic gonadotropin (rHCG) (Ovitrelle 250 mcg, Merck Serono, IL) when two to three leading follicles (18-19 mm in diameter) were demonstrated on ultrasound. Oocyte collection was performed 35-36 h later (Lahav-Baratz et al., 2004) .
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