Question

BELOW IS A METHOD. WHAT PROTOCOL (ALSO BELOW: A-F) MATCHES THIS METHOD?

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Method 5: Controlled ovarian stimulation and oocyte recovery was performed. Briefly, cycling females were subjected to follicular stimulation using twice-daily intramuscular injections of recombinant human FSH as well as concurrent treatment with Antide, a GnRH antagonist, for days 8-9. Unless indicated otherwise, all reagents were from Sigma-Aldrich and all hormones were from Ares Advanced Technologies Inc. Females received recombinant human luteinizing hormone on days 7-9 and recombinant human chorionic gonadotrophin (hCG) on day 10 primary culture of fibroblasts was established from an adult rhesus macaque male. Briefly, a small skin biopsy was surgically derived, washed in Cat2- and Mg+2 free Dulbecco PBS (Invitrogen) and minced into pieces before incubation in Dulbecco Modified Eagle's Medium (DMEM, Invitrogen) containing 1 mg m1-1 collagenase IV (invitrogen) at 37°C in 5% CO2 for 40 min. Tissue pieces were then vortexed, washed, seeded into 75 cm cell culture flasks (Corning) containing DMEM supplemented with 100 IU ml-1 penicillin, 100 mgml1 streptomycin (Invitrogen), 10% FBS (DMEM/FBS culture media) and cultured at 37°C in 5% CO2 until reaching confluency. Fibroblasts were then disaggregated with trypsin treatment and frozen down in aliquots of 13 106 cells in medium containing 10% dimethyl sulphoxide (DMSO). Fibroblasts were subsequently thawed, plated onto 4-well dishes (Nunc) and cultured under standard conditions until reaching 50-90% confluency. Cells were then synchronized in the GO/Gl phase of the cell cycle by culturing in DMEM medium with 0.5% FBS for 4 days before SCNT. Cumulus oocyte complexes were collected from anaesthetized animals by laparoscopic follicular aspiration (28-29 h after hCG) and placed in TALP/HEPES medium (modified Tyrode solution with albumin, lactate and pyruvate) containing 0.3% BSA(TH3) at 37°C, Oocytes were stripped of cumulus cells by mechanical pipetting after brief exposure (

Answer 1

Answer. Protocol A matches the method describe in this. As they enucleated the oocyte after M-II maturation by removing spindle then they fuse it with fibroblast [ somatic cell] and fertilization occurs that result in embryo formation and reaches the blastocyst stage.