Birth of healthy offspring following ICSI in in vitro-matured common marmoset (Callithrix jacchus) oocytes

PLoS One. 2014 Apr 21;9(4):e95560. doi: 10.1371/journal.pone.0095560. eCollection 2014.

Abstract

Intracytoplasmic sperm injection (ICSI), an important method used to treat male subfertility, is applied in the transgenic technology of sperm-mediated gene transfer. However, no study has described successful generation of offspring using ICSI in the common marmoset, a small non-human primate used as a model for biomedical translational research. In this study, we investigated blastocyst development and the subsequent live offspring stages of marmoset oocytes matured in vitro and fertilized by ICSI. To investigate the optimal timing of performing ICSI, corrected immature oocytes were matured in vitro and ICSI was performed at various time points (1-2 h, 2-4 h, 4-6 h, 6-8 h, and 8-10 h after extrusion of the first polar body (PB)). Matured oocytes were then divided randomly into two groups: one was used for in vitro fertilization (IVF) and the other for ICSI. To investigate in vivo development of embryos followed by ICSI, 6-cell- to 8-cell-stage embryos and blastocysts were nonsurgically transferred into recipient marmosets. Although no significant differences were observed in the fertilization rate of blastocysts among ICSI timing after the first PB extrusion, the blastocyst rate at 1-2 h was lowest among groups at 2-4 h, 4-6 h, 6-8 h, and 8-10 h. Comparing ICSI to IVF, the fertilization rates obtained in ICSI were higher than in IVF (p>0.05). No significant difference was noted in the cleaved blastocyst rate between ICSI and IVF. Following the transfer of 37 ICSI blastocysts, 4 of 20 recipients became pregnant, while with the transfer of 21 6-cell- to 8-cell-stage ICSI embryos, 3 of 8 recipients became pregnant. Four healthy offspring were produced and grew normally. These are the first marmoset offspring produced by ICSI, making it an effective fertilization method for marmosets.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Newborn
  • Blastocyst / cytology
  • Callithrix / embryology*
  • Cell Differentiation*
  • Embryo Transfer
  • Embryo, Mammalian / cytology
  • Embryo, Mammalian / metabolism
  • Embryonic Development
  • Female
  • Genetic Markers
  • Humans
  • Male
  • Microsatellite Repeats / genetics
  • Oocytes / metabolism*
  • Polar Bodies / cytology
  • Pregnancy
  • Sperm Injections, Intracytoplasmic / methods*

Substances

  • Genetic Markers

Grants and funding

A part of this study was the result of “Highly Creative Animal Model Development for Brain Sciences” and “Construction of System for Spread of Primate Model Animals” supported by the Strategic Research Program for Brain Science, Grant-in-Aid for Scientific Research A from the Ministry of Education, Culture, Sports, Science and Technology (MEXT; http://www.mext.go.jp/english/), Japan, PRESTO of the Japan Science and Technology Agency to E.S., Funding Program for World-Leading Innovative R&D on Science and Technology (FIRST) program “Strategic Exploitation of Neuro-Genetics for Emergence of the Mind” from the Cabinet Office (http://www.brain.riken.jp/first-okano/en/index.html), government of Japan to E.S. and H.O., and the Global COE Program for Education and Research Center for Metabolomic Systems Biology from the MEXT to M.S. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.