Human embryos cryopreserved at the blastocyst stage were thawed by a two-step rapid thawing protocol using Quinn's Advantage Thaw Kit (CooperSurgical, Trumbull, CT) as previously described35 ,36 . In brief, either cryostraws or vials were removed from the liquid nitrogen and exposed to air before incubating in a 37 °C water bath. Once thawed, embryos were transferred to a 0.5 mol l−1sucrose solution for 10 min followed by a 0.2 mol l−sucrose solution for an additional 10 min. The embryos were then washed in Quinn’s Advantage Medium with Hepes (CooperSurgical) plus 5% serum protein substitute (CooperSurgical) and each transferred to a 25 µl microdrop of either Quinn’s advantage cleavage medium (CooperSurgical) or Quinn’s advantage cleavage medium (CooperSurgical) supplemented with 10% serum protein substitute under mineral oil (Sigma, St Louis, MO). The embryos were cultured at 37 °C with 6% CO2, 5% O2 and 89% N2 under standard human embryo culture conditions in accordance with current clinical IVF practice. Embryos used in this study were days post fertilization (DPF) 5–6.
Quinn s advantage medium with hepes
Manufactured by CooperSurgical
Quinn's Advantage Medium with Hepes is a culture medium designed for the in vitro culture of human embryos. The medium contains the essential nutrients and growth factors required to support embryo development.
Automatically generated - may contain errors
Lab products found in correlation
Serum protein substitute, by CooperSurgical (2 mentions)
Serum protein substitute, by Merck Group (2 mentions)
Quinn s advantage thaw kit, by CooperSurgical (2 mentions)
Quinn s advantage cleavage medium, by CooperSurgical (2 mentions)
Microcentrifuge tube, by Eppendorf (1 mentions)
Dulbecco s phosphate buffered saline dpbs, by Thermo Fisher Scientific (1 mentions)
Polyvinyl alcohol (pva), by Merck Group (1 mentions)
Inverted microscope, by Leica (1 mentions)
3 protocols using quinn s advantage medium with hepes
1
Rapid Thawing and Culture of Human Blastocysts
2
Rapid Thawing and Culture of Human Blastocysts
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Human embryos cryopreserved at the blastocyst stage were thawed by a two-step rapid thawing protocol using Quinn's Advantage Thaw Kit (CooperSurgical, Trumbull, CT) as previously described35 ,36 . In brief, either cryostraws or vials were removed from the liquid nitrogen and exposed to air before incubating in a 37 °C water bath. Once thawed, embryos were transferred to a 0.5 mol l−1sucrose solution for 10 min followed by a 0.2 mol l−sucrose solution for an additional 10 min. The embryos were then washed in Quinn’s Advantage Medium with Hepes (CooperSurgical) plus 5% serum protein substitute (CooperSurgical) and each transferred to a 25 µl microdrop of either Quinn’s advantage cleavage medium (CooperSurgical) or Quinn’s advantage cleavage medium (CooperSurgical) supplemented with 10% serum protein substitute under mineral oil (Sigma, St Louis, MO). The embryos were cultured at 37 °C with 6% CO2, 5% O2 and 89% N2 under standard human embryo culture conditions in accordance with current clinical IVF practice. Embryos used in this study were days post fertilization (DPF) 5–6.
3
Biopsying Blastocysts for Cytogenetic Analysis
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The biopsy was performed on the heated stage of an inverted microscope (Leica Microsystems, Germany) equipped with a micromanipulation system (TransferMan®4m, CellTram®4m Air/ Oil, Eppendorf, Hamburg, Germany). The biopsy took place in buffered human tubal fluid (Quinn's Advantage™ Medium with HEPES, Cooper Surgical) and the penetration of the biopsy pipette (type MBB-FP-SM-30, Cooper Surgical) in the blastocoel for aspiration of the ICM was achieved with the assistance of a holding pipette (type MPH-MED-30, Cooper Surgical) and an OCTAX NaviLase system (Vitrolife). It has previously been shown that ICM and TE cells can be reliably separated by biopsy (Capalbo et al. 2013) . After collection of the ICM, a TE biopsy of 5-10 cells was performed on the opposite side of the ICM to avoid contamination of the TE sample. Both samples were separately washed in Dulbecco's phosphate-buffered saline (DPBS, Thermo Fisher Scientific, MA, USA) containing 0.1% polyvinyl alcohol (Sigma Aldrich, Missouri, USA) and were subsequently transferred in 0.5 ml Eppendorf microcentrifuge tubes containing 2.5 µl DPBS. Samples were kept on ice during the procedure and were then stored at -20°C for up to 7 days, until the cytogenetic analysis.
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