Production of Live Offspring from Vitrified-Warmed Oocytes Collected at Metaphase I Stage

PLoS One. 2016 Jun 22;11(6):e0157785. doi: 10.1371/journal.pone.0157785. eCollection 2016.

Abstract

Vitrification of matured oocytes is widely adopted in human clinics and animal research laboratories. Cryopreservation of immature oocytes, particularly those at metaphase I (MI), remains a challenge. In the present work, mouse MI oocytes denuded of cumulus cells were vitrified and warmed (V/W) either prior to (V/W-BEFORE-IVM, n = 562) or after (V/W-AFTER-IVM, n = 664) in vitro maturation (IVM). Derivative metaphase II (MII) oocytes were then used for intracytoplasmic sperm injection (ICSI). In the control groups, in vivo matured MII oocytes were used freshly (FRESH-MII, n = 517) or after V/W (MII-V/W, n = 617). In vitro and in vivo developmental competencies were compared among groups. Satisfactory blastocyst rates were achieved in V/W-BEFORE-IVM (27.5%) and V/W-AFTER-IVM (32.4%) groups, albeit as expected still lower than those from fresh-MII (56.1%) or MII-V/W (45.6%) oocytes. Similarly, the term development rates from V/W-BEFORE-IVM and V/W-AFTER-IVM were 12.4% and 16.7% respectively, acceptable but lower than those of the fresh-MII (41.2%) and MII-V/W (23.3%) groups. These data demonstrate that oocytes collected at MI stage are amenable to V/W, which can be performed before or after IVM with acceptable development rates including production of healthy pups. These findings provide useful knowledge to researchers and clinical practitioners for preservation and use of the otherwise discarded MI oocytes.

MeSH terms

  • Animals
  • Animals, Newborn
  • Cell Survival
  • Embryo Transfer
  • Embryonic Development
  • Female
  • In Vitro Oocyte Maturation Techniques
  • Male
  • Metaphase*
  • Mice, Inbred C57BL
  • Oocytes / cytology*
  • Specimen Handling / methods*
  • Sperm Injections, Intracytoplasmic
  • Vitrification*

Grants and funding

This study was supported by grants from Yupintan Traditional Chinese Medicine Foundation (Kaohsiung, Taiwan, R.O.C.) and the National Taiwan University, Taiwan, R.O.C., Grant # NTU-CDG-105R7885 to LYS and 103R4000 to WFC. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.