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.
Serum protein substitute
Manufactured by Merck Group
Serum protein substitute is a laboratory product that serves as a replacement for serum proteins. Its core function is to provide a source of proteins and other biomolecules that are typically found in serum, which can be used in various experimental and analytical procedures.
Automatically generated - may contain errors
2 protocols using serum protein substitute
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.
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