Florida Institute for Reproductive Medicine

Jacksonville, FL, United States

Florida Institute for Reproductive Medicine

Jacksonville, FL, United States
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Yang D.,Florida Institute for Reproductive Medicine | Winslow K.L.,Florida Institute for Reproductive Medicine | Nguyen K.,Florida Institute for Reproductive Medicine | Duffy D.,Florida Institute for Reproductive Medicine | And 2 more authors.
Journal of Experimental and Clinical Assisted Reproduction | Year: 2010

Background: This study examined the primary effect of selected cryoprotective agents (CPAs) on the meiotic spindles of human oocytes during cooling. Methods: Fresh metaphase II oocytes (n=26) donated from patients undergoing IVF treatment were analyzed via Polscope. In experiment one, 16 oocytes with visible spindle at 37°C were cooled to 20°C and rewarmed to 37°C to test the spindle response to cooling. They were then cooled to 20°C, 10°C, 0°C and rewarmed to 37°C after having been equilibrated with 1.5 M 1,2-propanediol (PROH), 1.5 M dimethyl sulfoxide (DMSO), 1.5 M ethylene glycol (EG) or 10 μM taxol at 37°C. In experiment two, 10 oocytes without visible spindles at 37°C were cooled to 20°C and then equilibrated with PROH, EG and taxol at 20°C. Spindle images were recorded at each temperature. Results: Meiotic spindles remained visible or became more distinct during cooling to 20°C, 10°C and 0°C when equilibrated with PROH, EG, DMSO and Taxol. Without these agents, meiotic spindles of the same oocytes disappeared after cooling to 20°C. Conclusion: The primary effect of PROH, EG and DMSO on the meiotic spindle is to stabilize and protect it against low temperature disassembly. A higher equilibration temperature (≥33°C) for oocyte freezing is recommended. © 2010 Yang et al.


PubMed | Florida Institute for Reproductive Medicine
Type: | Journal: Journal of experimental & clinical assisted reproduction | Year: 2010

This study examined the primary effect of selected cryoprotective agents (CPAs) on the meiotic spindles of human oocytes during cooling.Fresh metaphase II oocytes (n=26) donated from patients undergoing IVF treatment were analyzed via Polscope. In experiment one, 16 oocytes with visible spindle at 37C were cooled to 20C and rewarmed to 37C to test the spindle response to cooling. They were then cooled to 20C, 10C, 0C and rewarmed to 37C after having been equilibrated with 1.5 M 1,2-propanediol (PROH), 1.5 M dimethyl sulfoxide (DMSO), 1.5 M ethylene glycol (EG) or 10 M taxol at 37C. In experiment two, 10 oocytes without visible spindles at 37C were cooled to 20C and then equilibrated with PROH, EG and taxol at 20C. Spindle images were recorded at each temperature.Meiotic spindles remained visible or became more distinct during cooling to 20C, 10C and 0C when equilibrated with PROH, EG, DMSO and Taxol. Without these agents, meiotic spindles of the same oocytes disappeared after cooling to 20C.The primary effect of PROH, EG and DMSO on the meiotic spindle is to stabilize and protect it against low temperature disassembly. A higher equilibration temperature (33C) for oocyte freezing is recommended.

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