Multiparameter assessment of mouse oogenesis during follicular growth in vitro.
Comparison of oocyte development within the follicle in vitro and in vivo has a major impact on research into ovarian physiology and clinical practice. Despite major differences in ovarian physiology between rodents and humans, mice provide a useful model for studies of the endocrine and paracrine mechanisms controlling follicular development. In this study, early preantral follicles were isolated from 12-day-old mice and cultured individually in microdrops under oil during 6, 9 or 12 days. Taking into account previous observations, several oocyte criteria (diameter, chromatin configuration, transcriptional activity, intracytoplasmic calcium signalling and ability to undergo meiosis) were assessed to check that the development pattern of oocytes during follicle growth in vitro was similar to that already observed for oocytes developing in vivo, and that they reached the fertilizable oocyte stage. Results indicate that, during the 12-day-culture period, the oocytes grew until 74.3 +/- 4.2 microm, they became transcriptionally quiescent with a surrounded nucleolus (SN) chromatin organization, 50% of them exhibited regular calcium signals and 73.4% of them resumed meiosis. These data demonstrate that the protocol used generates oocytes with characteristics similar to oocytes allowed to mature fully in vivo and that it could be useful to set up the experimental culture of human ovarian follicles.
Pesty
A
A
INSERM, UF Biologie de la Reproduction, Paris, France. arlette.pesty@cea.fr
Miyara
F
F
Debey
P
P
Lefevre
B
B
Poirot
C
C
D000818:Animals; D020013:Calcium Signaling; D048429:Cell Size; D002843:Chromatin / Q000737:chemistry; D005260:Female; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D009865:Oocytes / Q000254:growth & development; D009866:Oogenesis / Q000502:physiology*; D006080:Ovarian Follicle / Q000254:growth & development* / Q000502:physiology; D046509:Tissue Culture Techniques; D014158:Transcription, Genetic
D002843:Chromatin