Applications of cryopreserved unfertilized mouse oocytes for in vitro fertilization

Cryobiology. 2013 Oct;67(2):188-92. doi: 10.1016/j.cryobiol.2013.06.011. Epub 2013 Jul 8.

Abstract

Since the first successful reports into oocyte freezing, many papers concerning the cryopreservation of mouse oocytes have been published. However, a simple and practical cryopreservation method for unfertilized C57BL/6 mouse oocytes, and an IVF system using these cryopreserved oocytes have yet to be established, in spite of the fact that C57BL/6 is the prevalent inbred strain and is used for large-scale knockout programs. In this study, unfertilized C57BL/6 mouse oocytes were cryopreserved via a simple vitrification method. After warming, IVF was performed using cryopreserved unfertilized oocytes and fresh sperm, cryopreserved unfertilized oocytes and cold-stored sperm, cryopreserved unfertilized oocytes and frozen sperm (C57BL/6 strain sperm), and cryopreserved unfertilized oocytes and frozen sperm derived from GEM strains (C57BL/6 background GEM strains). Nearly all of the cryopreserved oocytes were recovered, of which over 90% were morphologically normal. Those oocytes were then used for in vitro fertilization, resulting in 72-97% of oocytes developing into 2-cell embryos. A portion of the 2-cell embryos were transferred to recipients, resulting in live young being produced from 32-49% of the embryos. In summary, we established the simple and practical method of mouse oocyte vitrification with high survivability and developmental ability and the IVF using the vitrified-warmed oocytes with fresh, cold-stored or cryopreserved sperm with high fertility.

Keywords: Cold-stored sperm; Cryopreserved sperm; Fresh Sperm; In vitro fertilization; Mouse; Oocyte vitrification.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cryopreservation / methods*
  • Female
  • Fertility
  • Fertilization in Vitro / methods*
  • Freezing
  • Male
  • Mice / embryology
  • Mice / physiology*
  • Mice, Inbred C57BL
  • Oocytes / cytology*
  • Spermatozoa / cytology
  • Vitrification