Hyaluronidase

Manufactured by Irvine Scientific

Sourced in United States

Hyaluronidase is an enzyme that catalyzes the hydrolysis of hyaluronic acid, a major component of the extracellular matrix. It plays a role in tissue remodeling and can be used to facilitate the absorption and dispersion of injected fluids or other substances.

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Lab products found in correlation

14 protocols using hyaluronidase

Ovarian Stimulation and Oocyte Retrieval Protocol

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Ovarian stimulation was achieved using a gonadotropin-releasing hormone (GnRH) antagonist (Cetrorelix, Merck Serono, Berlin, Germany), recombinant follicle-stimulating hormone (FSH; Gonal-F, Merck Serono, Berne, Switzerland), and human menopausal gonadotropin (hMG, Livzon Pharmaceutical Group Inc., Zhuhai, China). Ultrasound-guided follicular puncture was performed 36 h after injecting human chorionic gonadotropin (hCG, Livzon Pharmaceutical Group Inc.). Harvested oocytes were denuded enzymatically with 2-hydroxyethyl (HEPES)-buffered medium containing hyaluronidase (Irvine scientific) and mechanically by pipetting with a commercial glass pipette (Origio). Morphologically normal spermatozoa were injected into metaphase-II (MII) mature oocytes.
Biochemical pregnancy was confirmed by positive β-human chorionic gonadotrophin (β-hCG) in the blood or urine 2 weeks after embryo transfer. Clinical pregnancy was confirmed 4–6 weeks after embryo transfer by ultrasonography based on the presence of a gestational sac and fetal heartbeat in the uterine cavity.

Chen W., Huang C., Li P., Liu F., Sun J., Zhu Z.J., Zhai J., Xu Y., Hong Y., Hu J.L., Peng Y.P., Zhang Z.B., Wu Y, & Li Z. (2022). Clinical benefits of a modified Cryopiece system for cryopreservation of rare ejaculated and testicular spermatozoa for ICSI. Asian Journal of Andrology, 24(5), 533-539.

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Optimizing ICSI Timing for Improved Outcomes

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The retrieved COC was incubated in fertilization medium (G‐IVF^TM; Vitrolife, Gothenburg, Sweden) until cumulus cell removal at 0 minutes (Group A) or 120 minutes (Group B) after oocyte retrieval by brief pipetting with 40 IU of hyaluronidase (Irvine Scientific, Santa Ana, CA, USA). Oocyte maturity was evaluated just before ICSI (not at the time of cumulus cells removal) in both the groups (Figure 1). ICSI was initiated between 2.5 and 4.0 hours after retrieval in both the groups (Figure 1). All ICSI procedures were completed within 5 hours after retrieval (Figure 1). Injected oocytes were cultured in human tubal fluid medium until fertilization verification on the next day.

Mizuno S., Ishikawa Y., Matsumoto H., Sato M., Ida M., Fukuda A, & Morimoto Y. (2018). The timing of cumulus cell removal for intracytoplasmic sperm injection influences the capability of embryonic development. Reproductive Medicine and Biology, 18(1), 111-117.

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Fertilization of Oocytes by ICSI

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Cumulus-oocyte complexes were cultured in fertilization medium (Life Global®, Cooper Surgical, Brussels, Belgium) at 37 °C, 5.7% CO₂, and 5% O₂. Fertilization was performed by insemination or by intracytoplasmic sperm injection (ICSI). Before ICSI, oocyte denudation was initiated by incubation in 80 IU/mL of hyaluronidase (Irvine Scientific, Santa Ana, CA, USA) followed by mechanical pipetting to remove the cumulus cells from the oocyte. ICSI procedures were performed using a Nikon Eclipse Ti microscope. The inseminated oocytes were inserted into the slides one day after oocyte retrieval when mechanical denudation from cumulus and corona radiata cells was completed.

Fruchter-Goldmeier Y., Kantor B., Ben-Meir A., Wainstock T., Erlich I., Levitas E., Shufaro Y., Sapir O, & Har-Vardi I. (2023). An artificial intelligence algorithm for automated blastocyst morphometric parameters demonstrates a positive association with implantation potential. Scientific Reports, 13, 14617.

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Controlled Ovarian Stimulation and IVF/ICSI Protocols

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All participants underwent a controlled ovarian stimulation that was processed based on the previous study (Wang et al., 2021 (link)). Oocytes were retrieved by transvaginal ultrasound 36–38 h after HCG administration. For IVF patients, oocytes got fertilized 3–4 h after oocyte retrieval, and degranulation occurred 4 h after fertilization. If the second polar body had not been observed until 6 h after fertilization, early rescue ICSI was performed after the patients signed a consent form. For ICSI patients, cumulus-oocyte complexes were exposed to 80 U/L hyaluronidase (Irvine Scientific, United States) followed by mechanical pipetting for degranulation. Nude oocytes were further cultured for another 1–2 h before spermatozoon injection. Generally, pronucleus (PN) assessments were performed 17–18 h after fertilization.

Wang M., Zhu L., Liu C., He H., Wang C., Xing C., Liu J., Yang L., Xi Q., Li Z, & Jin L. (2021). A Novel Assisted Oocyte Activation Method Improves Fertilization in Patients With Recurrent Fertilization Failure. Frontiers in Cell and Developmental Biology, 9, 672081.

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In Vitro Maturation and Fertilization of Oocytes

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Retrieved cumulus–oocyte complexes (COCs) were denuded of their cumulus and corona cells using mechanical and enzymatic treatment (80 IU ml–1 hyaluronidase, Irvine Scientific, Irvine, CA, USA) as described previously (21). The denuded oocytes were classified as either mature (MII) or immature (GV or MI) oocytes. Good quality immature oocytes were cultured in blastocyst medium (G-2 v5 Media, Vitrolife, Sweden) (22) supplemented with 75 mIU mL–1 FSH and 75 mIU mL–1 LH (Ferring) at 37 °C in an incubator under 5% CO2 and 95% air with high humidity. The maturity of the oocytes was assessed using a stereomicroscope (Olympus, Tokyo, Japan) 24 h after IVM. The matured oocytes were injected with spermatozoa. Fertilization was assessed 16–22 h after microinjection and the resulting zygotes were cultured in droplets of 20 µL of G-1 v5 Media (Vitrolife, Sweden) until the 8-cell stage.
Regarding the discarded embryos, on fertilization assessment day, the 1PN and 3PN zygotes and unfertilized oocytes were picked out from normally fertilized oocytes and were cultured in droplets of 20 µL of G-1™ v5 media (Vitrolife, Sweden), separately. Finally, grades A and B 8-cell cleaved embryos were selected for in vitro splitting.

OMIDI M., KHALILI* M.A., AGHA-RAHIMI A., NOTTOLA S.A., ANBARI F., FARAMARZI A, & PALMERINI M.G. (2021). Efficacy of the in vitro splitting of human preimplantation embryos from ART programs. Turkish Journal of Medical Sciences, 51(1), 68-75.

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ICSI Protocol for Mature Oocyte Retrieval

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Women were treated with GnRH agonist using either a short protocol or long protocol and with GnRH antagonist, according to each patient's ovarian response and medical history of IVF treatment. In some cycles, the minimal stimulation protocol was used, with a combination of low‐dose gonadotropin injection and clomiphene citrate. When a follicle reached a mean diameter of ≥18 mm, human chorionic gonadotropin (Aska Pharmaceutical) at a dose of 5000 or 10 000 IU was administered. Cumulus‐oocyte complexes were collected 35 hours after human chorionic gonadotropin administration using transvaginal ultrasound‐guided puncture of the follicles. Collected cumulus‐oocyte complexes were denuded by pipetting with 80 IU/mL hyaluronidase (Irvine Scientific). The denuded oocyte maturity was assessed by visualization of the first polar body under a stereoscopic microscope. Only MII oocytes were used for ICSI.

Harada Y., Maeda T., Fukunaga E., Shiba R., Okano S., Kinutani M, & Horiuchi T. (2019). Selection of high‐quality and viable blastocysts based on timing of morula compaction and blastocyst formation. Reproductive Medicine and Biology, 19(1), 58-64.

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Effects of RF Radiation on Mouse Embryos

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Ovulation was induced in mice by intraperitoneal (i.p.) injection of 10 IU of Pregnant Mare’s Serum Gonadotropin Folligon (PMSG; Intervet, Holland) followed by 10 IU of human chorionic gonadotropin (HCG; Organon, Holland) after 48 hr. In this study, one female mouse was mated with a male mouse overnight. The following morning, the mating was confirmed by inspection of the vaginal plug. Mice with visible copulation plugs were sacrificed by cervical dislocation, and then ovary burses were surgically removed and collected in Hams’F10 medium. Under stereomicroscope (Olympus.SZX16, Japan), zygotes were dissected out from the swollen ampulla and treated with hyaluronidase (80 IU/ml, Irvine Scientific, USA). The collected zygotes were transferred into 100 μl of G1 medium (Vitrolife, Sweden) and incubated for 5 hr in standard incubator. Next, 2-cell embryos were divided into two groups of control (n=150) and experimental (n=150). The latter group was exposed to RF radiation (30 min/day). Also, the embryos were cultured for a maximum of 4 days up to the blastocyst stage under liquid paraffin at 37°C/5% CO
₂. For recording any delay or abnormality during incubation, the morphology of the developing embryos was monitored daily under an inverted microscope (TE300; Nikon, Tokyo, Japan) equipped with heating stage.

Safian F., Khalili M.A., Khoradmehr A., Anbari F., Soltani S, & Halvaei I. (2016). Survival Assessment of Mouse Preimplantation Embryos After Exposure to Cell Phone Radiation. Journal of Reproduction & Infertility, 17(3), 138-143.

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Oocyte Injection with First vs Second Ejaculation

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The cumulus cells surrounding the retrieved oocytes were removed mechanically at 80 IU/ml hyaluronidase (Irvine Scientific, California, USA) preheated to 37°C, all MII oocytes were confirmed by one embryologist using an inverted microscope and then transferred into a droplet containing 30 μl of G-IVF PLUS™ (10136, Vitrolife) medium covered with mineral oil (Vitrolife, Goteborg, Sweden).Then, another embryologist randomly allocated the MII oocytes into two groups: the first ejaculation group, in which MII oocytes were injected with spermatozoons derived from the first ejaculation after a long abstinence period, and the second ejaculation group, in which MII oocytes were injected with spermatozoons derived from a short-interval second ejaculation. The injection procedure was performed by the same embryologist. The injected MII oocytes were transferred into G-1 PLUS™ (10128, Vitrolife), covered with mineral oil in a Primo Vision dish (9-well or 16-well, Vitrolife, Viby, Denmark) and cultured in a Primo Vision (Vitrolife, Budapest, Hungary) incubator at 37°C, 6% CO₂, and 5% O₂.

Li Y., Wang S., Li D., Huang Y., Liu H., Zhang X., Qin J., Mao X., Li Z., Chen L., Wei P., Shi W, & Xue L. (2023). Short-interval second ejaculation improves sperm quality, blastocyst formation in oligoasthenozoospermic males in ICSI cycles: a time-lapse sibling oocytes study. Frontiers in Endocrinology, 14, 1250663.

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Oocyte Denudation and ICSI Procedure

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Follicles were aspirated and the cumulus-oocyte complexes were washed in Global Total with HEPES medium (Life Global, Brussels, Belgium), oocytes were cultured in fertilization medium (Life Global, Brussels, Belgium) covered with mineral oil (Irvine Scientific, Santa Ana, CA, USA) for 3 h at 37 °C, 5.7% CO₂, and 5% O₂. Oocyte denudation was initiated by a 30-s incubation in 80 IU/mL of hyaluronidase (Irvine Scientific, Santa Ana, CA, USA), followed by washings in HEPES medium to remove enzyme residuals . Removal of cumulus cells from the oocyte was carried out by mechanical pipetting in Global Total medium containing HEPES. ICSI procedure was performed in the same medium at 400× magnification using a Nikon Eclipse Ti microscope.

Priel E., Priel T., Szaingurten-Solodkin I., Wainstock T., Perets Y., Zeadna A., Harlev A., Lunenfeld E., Levitas E, & Har-Vardi I. (2020). Zona pellucida shear modulus, a possible novel non-invasive method to assist in embryo selection during in-vitro fertilization treatment. Scientific Reports, 10, 14066.

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Oocyte Maturation and ICSI

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Follicles were punctured under ultrasound guidance 36 h after the administration of 10,000 IU human chorionic gonadotropin (hCG, Merck Serono, Switzerland). The corona-cumulus cells were removed by hyaluronidase (Irvine Scientific, USA), and then the meiotic status of human oocytes was assessed. Only MII-stage oocytes were fertilized by ICSI for the patients. Those GV-stage oocytes with a discernable germinal vesicle were donated and collected for this study.

Li Y., Liu H., Yu Q., Liu H., Huang T., Zhao S., Ma J, & Zhao H. (2019). Growth Hormone Promotes in vitro Maturation of Human Oocytes. Frontiers in Endocrinology, 10, 485.

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