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Animal Reproduction and Physiology
Asian-Australasian Journal of Animal Sciences 2007;20(8): 1190-1195.
DOI: https://doi.org/10.5713/ajas.2007.1190    Published online June 27, 2007.
Developmental Competence of Intrafollicular Oocytes Derived from Preantral Follicle Culture with Different Protocols after Parthenogenetic Activation
Jung Kyu Choi, Jae Hee Lee, Seung Tae Lee, Mun Hwan Choi, Seung Pyo Gong, Eun Ju Lee, Jeong Mook Lim*
Correspondence:  Jeong Mook Lim,
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 m in diameter) of F1 hybrid (B6CBAF1) mice were cultured singly for 216 h in modified -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
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