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The impact of vitrification on immature oocyte cell cycle and cytoskeletal integrity in a rat model

  • Gamete Biology
  • Published:
Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Purpose

To test the effects of varying vitrification protocols on the cell cycle status and chromosomal integrity in cumulus-enclosed GV stage rat oocytes.

Methods

Vitrified and thawed rat oocytes were labeled with fluorescent markers for chromatin, cell cycle activation, and f-actin and analyzed by conventional and laser scanning confocal microscopy.

Results

In all vitrification groups, significant alterations in cumulus cell connectivity, cell cycle status, and cytoplasmic actin integrity were observed following warming compared to fresh control oocytes. Based on the protein phosphorylation marker MPM-2, it is clear that warmed oocytes rapidly enter M-phase but are unable to maintain chromosome integrity as a result of multiple chromatin fusions. A prominent reduction in f-actin is evident in both the ooplasm and at the cortex of vitrified oocytes. Finally, an irreversible but irregular retraction of TZPs occurs on the majority of oocytes subjected to any of the vitrification protocols.

Conclusions

These findings draw attention to undesirable consequences of immature oocyte vitrification that compromise cell cycle status and chromatin and cytoskeleton integrity that may not be evident until after fertilization.

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Acknowledgments

We would like to extend our sincerest thanks to Kimball Pomeroy for statistical analysis and Darlene Limback for her continued technical support. Most of all, our special thanks and acknowledgment go to Sara Brown and Rebekah Lang without whom this study would not be possible. This project was supported by grants from the Eshe Fund (DFA), the National Institute of General Medical Sciences from the National Institutes of Health (P20 GM103318), and the National Institutes of Health (HD 20068 DFA).

Conflict of interest

The authors declare no conflicts of interest regarding this manuscript.

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Authors and Affiliations

  1. Department of Obstetrics and Gynecology, University of Kansas Medical Center, 3901 Rainbow Blvd, Kansas City, KS, 66160, USA

    S. Samuel Kim, Rachel Olsen, Dojun David Kim & David F. Albertini

  2. Department of Molecular and Integrative Physiology, University of Kansas Medical Center, Kansas City, KS, USA

    S. Samuel Kim, Rachel Olsen, Dojun David Kim & David F. Albertini

Authors
  1. S. Samuel Kim
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  2. Rachel Olsen
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  3. Dojun David Kim
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  4. David F. Albertini
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Correspondence to S. Samuel Kim.

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Capsule

A comparison of vitrification protocol variations on cumulus-enclosed rat GV oocytes has revealed dramatic effects on cell cycle status, somatic cell-oocyte interactions, and the stability of the actin cytoskeleton.

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Kim, S.S., Olsen, R., Kim, D.D. et al. The impact of vitrification on immature oocyte cell cycle and cytoskeletal integrity in a rat model. J Assist Reprod Genet 31, 739–747 (2014). https://doi.org/10.1007/s10815-014-0216-1

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  • DOI: https://doi.org/10.1007/s10815-014-0216-1

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