The patients received standard ovarian stimulation utilizing either a long or short protocol. A gonadotropin-releasing hormone analog downregulates (Triptorelin Acetate, Ferring AG, Switzerland), the patients were stimulated with recombinant FSH (Gonal-F; Serono, Switzerland). When three or more follicles reached 18 mm in diameter, 6000~10000 IU human chorionic gonadotropin (hCG) (Lizhu, China) was administered. Ultrasound guided vaginal puncture was conducted for oocyte retrieval within 34–36 h after hCG injection. The cumulus-oocyte complexes (COCs) were isolated and cultured in fertilization medium at 37°C with 6% CO2 for 2~4 h. After denudation of COCs, the oocytes with a GV structure were used for subsequent experiments. Prior to cryopreservation, the oocytes were microscopically assessed for their structure to identify superior oocytes of suitable dimensions, intact zona pellucida, and intact membrane.
Triptorelin acetate
Manufactured by Ferring
Sourced in Switzerland
Triptorelin acetate is a synthetic gonadotropin-releasing hormone (GnRH) agonist. It is used in the treatment of various medical conditions that require the regulation of reproductive hormone levels.
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8 protocols using triptorelin acetate
1
Oocyte Retrieval and Cryopreservation Protocol
2
Ultra-long GnRH Agonist Protocol for IVF
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Ultra-long GnRH agonist protocol has been described previously (7 (link)). Patients received a depot injection of triptorelin acetate (3.75mg; Ferring Pharmaceuticals, Kiel, Germany) 28~30 days before ovarian stimulation on 2~4 days of menstrual cycle and COS started 28~35 days after the last injection. When at least three dominant follicles reached the diameter of 18~20mm, recombinant hCG (250ug; Merck Serono, Coinsins, Switzerland) was injection intramuscularly. The oocytes aspiration was performed 36.5~37.5 hours after trigger.
3
Controlled Ovarian Stimulation Protocols
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Each patient accepted a controlled ovarian stimulation (COS) regimen such as long, ultralong, and microstimulation protocol according to their ovarian function and responses during previous IVF cycles or ovulation induction. The diameter of follicles was monitored every 2-3 days, and ovulation was induced with human chorionic gonadotropin (hCG; Choragon, Ferring, Switzerland, 5000-10,000 IU) alone or in combination with triptorelin acetate (Ferring, 0.2 mg) when at least two leading follicles grew up to 18 mm.
4
Ovarian Stimulation Protocols for IVF
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Different COS regimens, i.e. long, antagonist and others (minimal stimulation, natural, luteal phase stimulation and progestinprimed stimulation), were selected for ovarian stimulation according to patients' characteristics and responses during previous IVF cycles or ovulation induction. When the diameter of at least one leading follicle was 18 mm, ovulation was induced with 5000-10,000 IU of human chorionic gonadotropin (hCG; Choragon, Ferring, Switzerland), alone or in combination with 0.2 mg triptorelin acetate (Ferring). Transvaginal ultrasound-guided oocyte retrieval was performed at 36 h after hCG administration.
5
Evaluating Chemotherapy Protection in Mice
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Female ICR mice, weighing 30–35 g, were randomly assigned to four groups: The vehicle, PXL, GnRHa and PXL+GnRHa groups. Based on our pre-examination and a previous study (27 (link)), the estrous cycle of ICR mice was 5 days on average. Therefore, GnRHa (1 mg/kg, triptorelin acetate; Ferring Pharmaceuticals) or normal saline was administered intraperitoneally to mice prior to chemotherapy for 5 days. Next, animals received a single dose of PXL (30 mg/kg; Pfizer, Inc.) or vehicle intraperitoneally. One quarter of the mice in each group were assessed 24 h after chemotherapy. The rest of the mice were continuously administered GnRHa for another estrous cycle (5 days) following chemotherapy, and were assessed on days 6, 11 and 16 following chemotherapy (Fig. 1A ). Another set of mice was administerd 30 mg/kg PXL every 3 days, for a total of 3 doses. GnRHa (1 mg/kg) was also administered prior to, during and following chemotherapy for a total of 16 days, and mice were then assessed on days 1, 6, 11 and 16 following chemotherapy (Fig. 1B ). During the experiment, one mouse in the GnRHa group suffered from weight loss and was euthanized before the endpoint of our experiment.
6
Ovulation Induction Protocols for IVF
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Different COS regimens (i.e., the long, antagonist, and others (minimal-stimulation, natural, luteal phase stimulation, and progestin-primed stimulation)) were used according to the patients' characteristics and responses during previous IVF cycles or ovulation induction. When the diameter of at least one leading follicle was 18 mm, ovulation was induced with 5000–10,000 IU of human chorionic gonadotropin (hCG; Choragon, Ferring, Switzerland), alone or in combination with 0.2 mg triptorelin acetate (Ferring). Transvaginal ultrasound-guided oocyte retrieval was performed at 36 h after hCG administration.
7
GnRH-a Induced Ovarian Stimulation Protocol
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A total of 310 patients were recruited in the long-acting protocol (LAP) group induced by a combination of GnRH-a incorporated with Follicular stimulating hormone (FSH) and Human menopausal gonadotropin (hMG). Women with a regular cycle on day 21 in the mid-luteal phase started with the administration of a single intramuscular dose of 0.1-1.2 mg long-acting decapeptyl® (Triptorelin acetate; Ferring) GnRH-a injection. A complete down-regulation of the pituitary had done when the serum LH level < 2 IU/ml and serum E2 level < 30 pg/mL was achieved. The transvaginal ultrasound scan (TVS) revealed a less than 5mm endometrium thickness, con rming complete pituitary suppression. On cycle day 2, exogenous gonadotropin rFSH (5.5µg; Gonal-F™, Merk Serono) and menotropins hMG (LG™ life sciences, Korea) administration were commenced at doses ranging between 75-220 IU/day and 350-450 IU/day depending upon body weight, age, and follicular size of the patients. A further regular dosage of rFSH and hMG was calculated based on ovarian stimulation monitored through transvaginal ultrasound scan (TVS) and serum E2 levels. The folliculogenesis was consecutively observed through TVS and by measuring the serum E2, LH, and progesterone ratio from the 8th day to the day of Human chorionic gonadotropin (hCG) injection (Pregnyl®, Organon), which is around 14 days post-GnRH-a administration.
8
Triptorelin-Guided Controlled Ovarian Stimulation
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A total of 230 patients in the short-acting protocol group have got a 0.1 mg daily intramuscular dose of decapeptyl® (Triptorelin acetate; Ferring) started from day 3rd of the menstrual cycle and continued till the day of hCG. Controlled ovarian stimulation started from 2nd day of menstrual cycle at the dose of 100-220 IU of rFSH (5.5µg; Gonal-F™, Merk Serono) and 200-450 IU of hMG (LG™ life sciences, Korea) respectively. The daily dosage was adjusted in accordance with ovarian response.
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