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Developmental Competence of Intrafollicular Oocytes Derived from Preantral Follicle Culture with Different Protocols after Parthenogenetic Activation

  • Choi, Jung Kyu (Department of Food and Animal Biotechnology, Seoul National University) ;
  • Lee, Jae Hee (Department of Food and Animal Biotechnology, Seoul National University) ;
  • Lee, Seung Tae (Department of Food and Animal Biotechnology, Seoul National University) ;
  • Choi, Mun Hwan (Department of Food and Animal Biotechnology, Seoul National University) ;
  • Gong, Seung Pyo (Department of Food and Animal Biotechnology, Seoul National University) ;
  • Lee, Eun Ju (Department of Food and Animal Biotechnology, Seoul National University) ;
  • Lim, Jeong Mook (Department of Food and Animal Biotechnology, Seoul National University)
  • Received : 2006.10.19
  • Accepted : 2007.04.02
  • Published : 2007.08.01

Abstract

This study was conducted to improve efficiency of a follicle culture system without reducing developmental competence of intrafollicular oocytes. Preantral follicles (100 to $125{\mu}m$ in diameter) of F1 hybrid (B6CBAF1) mice were cultured singly for 216 h in modified ${\alpha}$-MEM-glutamax medium, to which 2.5 IU/ml hCG and epidermal growth factor was added 16 h prior to the end of culture. Medium change was either performed three times (54 h interval), twice (72 h interval), once (108 h interval), or not at all (216 h interval). Maturation (progression to the metaphase II stage) of intrafollicular oocytes was detected from 4 days after culture in the three-times change treatment, while all treatments yielded mature oocytes from day 5 of culture. Compared with the three-times change, decreasing the change frequency to once did not reduce the capacity to begin maturation (germinal vesicle breakdown of 82 to 86%), to mature (78 to 79%) and to develop into blastocysts after parthenogenetic activation (29 to 32%). Morphological parameters were similar among these treatments. Except for the no medium change treatment, similar colony-forming activity of inner cell mass cells after culturing of blastocysts in leukemia inhibitory factor-containing medium was detected, while the morphology of the colony-forming cells deteriorated in the change-once treatment compared with the change twice or three-times. In conclusion, the efficiency of the preantral follicle culture system could be improved by reducing frequency of medium change up to a 72 h interval (three times in total 216 h culture) without decreasing developmental competence of oocytes.

Keywords

Mouse;Secondary Follicle;Intrafollicular Oocyte;Maturation;Parthenogenesis;Blastocyst Formation

Acknowledgement

Supported by : Ministry of Science and Technology

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