IVF is a technique by which eggs retrieved from females are fertilized in vitro with spermatozoa. IVF was performed as described previously12 . Females were superovulated by injecting 10 IU of pregnant mare serum gonadotropin (ASKA Pharmaceutical Co., Ltd., Tokyo, Japan; to bring the oocytes to maturity), followed by 10 IU of human chorionic gonadotropin (ASKA Pharmaceutical Co.; to release the oocytes) 48 h later. Ovulated oocytes were collected from the oviducts 14 h after human chorionic gonadotropin injection. Cumulus-enclosed oocytes were placed in 200-µL drops of TYH medium17 (link) and covered with paraffin oil (Nacalai Tesque, Kyoto, Japan). Spermatozoa were collected by mechanically dissecting the cauda epididymites and were placed in 200-µL drops of TYH medium. After 2 h of incubation (in this duration, sperm gains the ability to fertilize), the sperm suspension in TYH was added to the TYH drop containing eggs at a concentration of 100 sperm/μL. After 2 h incubation at 37 °C under 6% CO2 in air (in this duration, spermatozoa fertilizes with oocytes), cumulus cells were dispersed by brief treatment with hyaluronidase (Type-IS, 150 U/mL, Sigma, St. Louis, MO, USA). Three hours after the dispersion of cumulus cells, the number of pronuclei was counted to check the fertilization (2 pronuclear embryos are the fertilized embryos).
Pregnant mare serum gonadotropin
Manufactured by Aska Pharmaceutical
Sourced in Japan
Pregnant mare serum gonadotropin is a hormone extracted from the serum of pregnant mares. It contains a mixture of follicle-stimulating hormone (FSH) and luteinizing hormone (LH), which play a role in regulating reproductive processes.
Automatically generated - may contain errors
Lab products found in correlation
Human chorionic gonadotropin (hcg), by Aska Pharmaceutical (10 mentions)
Hyaluronidase type 1 s, by Merck Group (2 mentions)
Bovine testicular hyaluronidase, by Merck Group (2 mentions)
Human chorionic gonadotropin (hcg), by Merck Group (2 mentions)
Paraffin oil, by Nacalai Tesque (1 mentions)
Hepes czb medium, by Merck Group (1 mentions)
Paraffin liquid, by Nacalai Tesque (1 mentions)
Dylight 488, by Thermo Fisher Scientific (1 mentions)
Bz x810 microscope, by Keyence (1 mentions)
11 protocols using pregnant mare serum gonadotropin
1
In Vitro Fertilization (IVF) Protocol
2
Isolation of Mature Mouse Oocytes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mature oocytes were collected from the oviducts of 8- to 10-week-old female mice that had been induced to superovulate with 5 IU pregnant mare serum gonadotropin (ASKA Pharmaceutical, Tokyo, Japan) followed by 5 IU human chorionic gonadotropin (hCG) (ASKA Pharmaceutical) 48 h later. Cumulus–oocyte complexes (COCs) were collected from the oviducts ~16 h after hCG injection. To collect small denuded MII oocytes, COCs were placed in HEPES-buffered CZB medium and treated with 0.1% bovine testicular hyaluronidase (Sigma-Aldrich, St Louis, MO, USA). After several minutes, the COCs were washed twice and placed into a droplet of CZB medium for culture.
3
Superovulation and Embryo Collection Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Females were induced to superovulate by an intraperitoneal injection of 150 IU/kg pregnant mare serum gonadotropin (ASKA Pharmaceutical Co. Ltd., Tokyo, Japan) followed by an injection of 75 IU/kg human chorionic gonadotropin (ASKA Pharmaceutical Co. Ltd.) 48 h later. Females were then mated with males of the same strain overnight. Pronuclear-stage embryos were collected from oviducts of females the day after mating, and they were kept in modified Krebs-Ringer bicarbonate (mKRB)25 (link) before electroporation.
4
Superovulation and Oocyte Collection in Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Female mice were induced to undergo superovulation via treatment with 5 IU pregnant mare serum gonadotropin (ASKA Pharmaceutical Co., Ltd., Tokyo, Japan) followed 48 hours later by 5 IU human chorionic gonadotropin (hCG) (ASKA Pharmaceutical Co., Ltd.). Sixteen hours after hCG treatment, cumulus–oocyte complexes (COCs) were collected from the oviducts as quickly as possible and placed into HEPES CZB medium containing 0.1% bovine testicular hyaluronidase (Sigma-Aldrich, St Louis, MO, USA). After several minutes, oocytes were washed and placed into a droplet of CZB medium for culture.
5
Mouse In Vitro Fertilization Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
IVF was performed as described previously14 . Briefly, females were superovulated by injecting 10 IU of pregnant mare serum gonadotropin (ASKA Pharmaceutical Co., Ltd., Tokyo, Japan) to bring the oocytes to maturity. This was followed by administering 10 IU of human chorionic gonadotropin (ASKA Pharmaceutical Co.) 48 h later to release the oocytes. Ovulated oocytes were collected from the oviducts 14 h after injection. Cumulus-enclosed oocytes were placed in 200 µL drops (2 drops) of TYH medium33 (link) and covered with liquid paraffin (Nacalai Tesque, Kyoto, Japan). Spermatozoa were collected by mechanically dissecting the cauda epididymites and were placed in 200 µL drops of TYH medium. After 2 h of incubation, spermatozoa gained the ability to fertilize, and the sperm suspension was added to the TYH drops containing eggs at a concentration of 100 sperm/μL and incubated for 2 h at 37 °C under 6% CO2 in air to allow spermatozoa to fertilize the oocytes. Cumulus cells were dispersed by brief treatment with hyaluronidase (Type-IS, 150 U/mL, Sigma, St. Louis, MO, USA). Three hours after the dispersion of cumulus cells, the number of pronuclei was counted to check for fertilization.
6
Superovulation and Oocyte Harvesting in Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
B6D2F1/Crlj female mice 8–16 weeks old (Charles River Japan Inc., Yokohama, Japan) were used as oocyte donors. Animals were housed in plastic cages in a specific pathogen-free barrier facility that was air-conditioned (temperature 24±1°C, humidity 40±10%) and light-controlled (lights on from 08:00 to 20:00). All animal care and procedures performed in this study conformed to the Guidelines for Animal Experiments of Kyoto University, and were approved by the Animal Research Committee of Kyoto University.
Females were induced to superovulate by an intraperitoneal injection of 5 IU of pregnant mare serum gonadotropin (ASKA Pharmaceutical. Co., Ltd., Tokyo, Japan) followed by an injection of 5 IU of human chorionic gonadotropin (ASKA Pharmaceutical. Co., Ltd.) 48 h later. Cumulus-oocyte complexes were collected from oviducts 13 to 15 h after the injection of human chorionic gonadotropin, and oocytes were freed from cumulus cells by treatment with 0.1% hyaluronidase in H-CZB medium for 5 min. Oocytes were rinsed in fresh H-CZB medium and kept at room temperature before sperm injection.
Females were induced to superovulate by an intraperitoneal injection of 5 IU of pregnant mare serum gonadotropin (ASKA Pharmaceutical. Co., Ltd., Tokyo, Japan) followed by an injection of 5 IU of human chorionic gonadotropin (ASKA Pharmaceutical. Co., Ltd.) 48 h later. Cumulus-oocyte complexes were collected from oviducts 13 to 15 h after the injection of human chorionic gonadotropin, and oocytes were freed from cumulus cells by treatment with 0.1% hyaluronidase in H-CZB medium for 5 min. Oocytes were rinsed in fresh H-CZB medium and kept at room temperature before sperm injection.
7
Blastocyst Collection from Superovulated Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For blastocyst collection, 8-week-old C57BL/6N female mice (Crea Inc., Tokyo, Japan) were superovulated with an intraperitoneal (i.p.) injection of 5 IU of pregnant mare serum gonadotropin (Aska Pharmaceutical Co. Ltd., Tokyo, Japan) followed by an i.p. injection of 5 IU of human chorionic gonadotropin (hCG; Aska Pharmaceutical Co. Ltd.) 48 h later. These superovulated female mice were mated with C57BL/6N(+/Y) or Ftx-deficient mice (−/Y) and blastocysts were collected from the uterus on 3.5 dpc from around 92–100 h post-hCG.
8
Superovulation and Mating Procedure
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Pregnant mare serum gonadotropin (5 units; ASKA Pharmaceutical) or Center for Animal Resources and Development (CARD) HyperOva (0.1 mL; Kyudo) was injected into the abdominal cavity of B6D2F1 females, followed by natural mating with mutant male mice 12 h after human coagulating gland (hCG; 5 units, ASKA Pharmaceutical) injection. After 6 h to 9 h of mating, we observed the collected oocytes.
9
Fc-fusion Protein Binding to Zona-free Hamster Eggs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Sexually mature female Syrian golden hamsters (Japan SLC Inc.) (approved by the Animal Care and Use Committee of the Research Institute for Microbial Diseases, Osaka University #28-4-2) were superovulated by peritoneal injection of pregnant mare serum gonadotropin and human coagulating gland (20 units for each; ASKA Pharmaceutical). Cumulus-oocyte complexes were extracted from the oviductal ampulla and treated with 1 mg/mL collagenase to remove the cumulus cells and zona pellucida, which yields zona-free eggs. These zona-free eggs were incubated with 200 nM Fc-fusion proteins in Biggers-Whitten-Whittingham medium (11 ) for 1 h and then stained with goat anti-human IgG Fc antibody DyLight 488 (Invitrogen) at a dilution of 1:50 for 1 h at 37 °C, 5% CO2. The eggs were imaged under a Keyence BZ-X810 microscope.
10
In Vitro Fertilization of Mouse Oocytes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Adult mice (>10 weeks of age) were used for in vitro fertilization (IVF). Briefly, spermatozoa were collected from the epididymides of adult males and incubated in droplets of human tubal fluid (HTF) medium for 1 h at 37 °C under 5% CO2 in humidified air. Cumulus–oocyte complexes (COCs) were collected from the oviducts of female mice that had been superovulated by injecting 7.5 IU of pregnant mare serum gonadotropin (ZENOAQ, Tokyo, Japan) and/or anti-inhibin serum [16 (link)] (a gift from Dr Gen Watanabe) followed by 7.5 IU of human chorionic gonadotropin (hCG; ASKA Pharmaceutical Co., Ltd., Tokyo, Japan) at a 48-h interval. Then, 15–17 h after the hCG injection, the COCs were isolated and incubated in HTF medium containing 0.125–1.25 mM glutathione (l-glutathione reduced, #G6013; Merck, Darmstadt, Germany) for 1 h before insemination. Next, preincubated spermatozoa were introduced to the COC-containing HTF droplets. At 5–6 h after insemination, fertilized zygotes were washed, transferred to droplets of potassium-enriched simplex optimized medium (KSOM) and cultured as above.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!