This study was approved by the Eulji University Institutional Animal Care and Use Committee (No. EUIACUC 17-14). The protocol for superovulated mice was described by Park et al. [25 ]. Briefly, 6- to 9-week-old female BDF mice were superovulated with intraperitoneal injections of 5 IU of pregnant mare’s serum gonadotropin (Prospec, Rehovot, Israel), and 48 hours later, the mice were injected with 5 IU of human chorionic gonadotropin (Prospec). The superovulated mice were then individually mated with a fertile male BDF mouse.
Pregnant mare s serum gonadotropin
Manufactured by ProSpec
Sourced in United States
Pregnant mare's serum gonadotropin is a hormone extracted from the blood of pregnant mares. It is used in laboratory settings to stimulate ovarian follicle development and ovulation in various animal species.
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Lab products found in correlation
3 protocols using pregnant mare s serum gonadotropin
1
Superovulation Protocol for Breeding Mice
2
CRISPR Genome Editing in Mouse Embryos
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Superovulation was induced in C57BL/6 female mice by injection of pregnant mare’s serum gonadotropin (ProSpec, NJ, USA) and human chorionic gonadotropin (ProSpec), followed by embryo collection on the following day. After 1, 2 h incubation, viable embryos with two pronuclei were selected, and microinjection was conducted using a micromanipulator (Eppendorf, Germany). Briefly, 50 ng/μL of Cas9 mRNAs, 10–20 ng/μL of each sgRNAs, and 20 ng/μL of each ssODNs were mixed and microinjected into pronucleus or cytoplasm of embryos. Embryos were cultured to the two-cell stage, followed by transfer into pseudopregnant females or culturing until the blastocyst stage for genotyping.
3
Porcine Oocyte In Vitro Maturation with SHH Protein
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Porcine ovaries were collected from a local slaughterhouse and transported to the laboratory in a sterile 0.9% NaCl solution. The COCs were recovered from ovarian follicles sized 3–6 mm by aspiration with an 18-gauge needle and a 10-mL syringe. Only COCs with a minimum of three cumulus cell layers were used for IVM. Before IVM, the COCs were classified as BCB− or BCB+, according to their coloration through BCB staining. Then, BCB− or BCB+ COCs were cultured with or without 0.5 µg/mL recombinant mouse SHH protein (SHH N-Terminus; 461-SH-025; R&D Systems, Minneapolis, MN, USA) during the entire period of IVM. The concentration of SHH protein was set according to a previous study [15 (link)]. The IVM medium was composed of tissue culture medium-199 supplemented with 10 ng/mL epidermal growth factor, 10 ng/mL β-mercaptoethanol, 0.57 mM cysteine, 10% porcine follicular fluid, 10 IU/mL human chorionic gonadotropin (Prospec, East Brunswick, NJ, USA), and 10 IU/mL pregnant mare’s serum gonadotropin (Prospec). The COCs were cultured in IVM medium under 5% CO2 at 38.5 °C for 22 h. They were then washed and cultured in hormone-free IVM medium for an additional 22 h.
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