Optimization of Cryoprotectant Loading into Murine and Human Oocytes Public Deposited

http://ir.library.oregonstate.edu/concern/articles/k643b3030

This is an author's peer-reviewed final manuscript, as accepted by the publisher. The published article is copyrighted by Elsevier and can be found at:  http://www.journals.elsevier.com/cryobiology/.

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  • Loading of cryoprotectants into oocytes is an important step of the cryopreservation process, in which the cells are exposed to potentially damaging osmotic stresses and chemical toxicity. Thus, we investigated the use of physics-based mathematical optimization to guide design of cryoprotectant loading methods for mouse and human oocytes. We first examined loading of 1.5 M dimethylsulfoxide (Me₂SO) into mouse oocytes at 23°C. Conventional one-step loading resulted in rates of fertilization (34%) and embryonic development (60%) that were significantly lower than those of untreated controls (95% and 94%, respectively). In contrast, the mathematically optimized two-step method yielded much higher rates of fertilization (85%) and development (87%). To examine the causes for oocyte damage, we performed experiments to separate the effects of cell shrinkage and Me₂SO exposure time, revealing that neither shrinkage nor Me₂SO exposure single-handedly impairs the fertilization and development rates. Thus, damage during one-step Me₂SO addition appears to result from interactions between the effects of Me₂SO toxicity and osmotic stress. We also investigated Me₂SO loading into mouse oocytes at 30°C. At this temperature, fertilization rates were again lower after one-step loading (8%) in comparison to mathematically optimized two-step loading (86%) and untreated controls (96%). Furthermore, our computer algorithm generated an effective strategy for reducing Me₂SO exposure time, using hypotonic diluents for cryoprotectant solutions. With this technique, 1.5 M Me₂SO was successfully loaded in only 2.5 min, with 92% fertilizability. Based on these promising results, we propose new methods to load cryoprotectants into human oocytes, designed using our mathematical optimization approach.
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  • Karlsson, J. O. M., Szurek, E. A., Higgins, A. Z., Lee, S. R., & Eroglu, A. (2014). Optimization of cryoprotectant loading into murine and human oocytes. Cryobiology, 68(1), 18-28. doi:10.1016/j.cryobiol.2013.11.002
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  • description.provenance : Made available in DSpace on 2014-08-04T21:18:32Z (GMT). No. of bitstreams: 1 HigginsAdamChemBioEnvEngOptimizationCryoprotectantLoading.pdf: 694088 bytes, checksum: 1b5c14cf945bba1c810732d262d5136a (MD5) Previous issue date: 2014-02
  • description.provenance : Approved for entry into archive by Deanne Bruner(deanne.bruner@oregonstate.edu) on 2014-08-04T21:18:32Z (GMT) No. of bitstreams: 1 HigginsAdamChemBioEnvEngOptimizationCryoprotectantLoading.pdf: 694088 bytes, checksum: 1b5c14cf945bba1c810732d262d5136a (MD5)
  • description.provenance : Submitted by Deanne Bruner (deanne.bruner@oregonstate.edu) on 2014-08-04T21:17:31Z No. of bitstreams: 1 HigginsAdamChemBioEnvEngOptimizationCryoprotectantLoading.pdf: 694088 bytes, checksum: 1b5c14cf945bba1c810732d262d5136a (MD5)

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