Asian-Australasian Journal of Animal Sciences

Asian Australasian Association of Animal Production Societies (아세아태평양축산학회)
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In vitro Culture Conditions for the Mouse Preantral Follicles Isolated by Enzyme Treatment

  • Kim, Dong-Hoon (Animal Biotechnology division, National Institute of Animal Science) ;
  • Seong, Hwan-Hoo (Animal Biotechnology division, National Institute of Animal Science) ;
  • Lee, Ho-Joon (Vincent Center for Reproductive Biology, Vincent Obstetrics and Gynecology Service, Harvard Medical School)
  • Received : 2007.09.01
  • Accepted : 2007.11.29
  • Published : 2008.04.01
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Abstract

In order to investigate the factors affecting the culture of mouse preantral follicles in vitro, we examined the effect of culture media, protein supplements, and culture period on their growth. The oocyte diameter (initial size: $55.6{\pm}2.5{\mu}m$) was progressively increased during culture, and the maximum size ($72.0{\pm}2.4{\mu}m$) was reached on day 10 of the in vitro culture. The chromatin configuration in the germinal vesicle (GV) oocyte progressively shifted from a non-surrounded nucleolus (NSN) to a surrounded nucleolus (SN). On day 10 of the culture, most of the oocytes progressed to the SN pattern. The survival and metaphase II rates of the oocytes in alpha-minimal essential medium (alpha-MEM) were significantly higher (p<0.05) than those in Waymouth and tissue culture medium (TCM)-199. As a protein source, fetal bovine serum (FBS) was more suitable for the culture of mouse preantral follicles as compared to human follicular fluid (hFF) and bovine serum albumin (BSA); the optimal concentration of FBS was 5%. These results suggest that in a culture of mouse preantral follicles, alpha-MEM and 5% FBS are an optimal medium and a protein source, respectively; further, the 10 days of culture is required for the complete growth of oocytes in this culture system.

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References

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