PMC2700691_01
Frozen oocytes
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Cook's oocyte freezing media (Cook Medical, Queensland, Australia) was used after warming the required quantity to room temperature, the active cryoprotectants being propanediol in increasing concentrations from 0.75 to 1.5 mol/l (solutions 1 and 2 - for 7min and 30 s, respectively) and then transferred to solution 3 containing 1.5 mol/l propanediol and 0.2 mol/l sucrose [Figure 1] for 5 min.
Thawed oocytes
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A survival check carried out under the inverted microscope revealed eight oocytes that survived the thawing process [Figure 2].
Holding egg
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Trumbull CT, USA) at 37 C and 5% CO2 for 2 h before injection. Subsequently, the oocytes were injected with frozen-thawed and prepared sample of husband's sperms. Intracytoplasmic sperm injection [Figures 3 and 4] was the preferred method for fertilization, owing to an expected zona hardening following oocyte cryopreservation. The injected oocytes were further cultured in cleavage media in a Trigas incubator until Pronucleus check 16-18h later.
Intracytoplasmic sperm injection
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Trumbull CT, USA) at 37 C and 5% CO2 for 2 h before injection. Subsequently, the oocytes were injected with frozen-thawed and prepared sample of husband's sperms. Intracytoplasmic sperm injection [Figures 3 and 4] was the preferred method for fertilization, owing to an expected zona hardening following oocyte cryopreservation. The injected oocytes were further cultured in cleavage media in a Trigas incubator until Pronucleus check 16-18h later.
Embryos
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Five embryos (two and three cells, grade II-III) [Figures 5 and 6] were observed and transferred on Day 2 using a Labotect catheter (Labotect GmbH, Gottinggen, Germany).
Embryos
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Five embryos (two and three cells, grade II-III) [Figures 5 and 6] were observed and transferred on Day 2 using a Labotect catheter (Labotect GmbH, Gottinggen, Germany).
At birth
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She delivered a healthy male baby weighing 2.54 kg [Figure 7].
At 3 months
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A general follow-up at 3 months showed normal development for age [Figure 8].