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Developmental Competence of Intrafollicular Oocytes Derived from Preantral Follicle Culture with Different Protocols after Parthenogenetic Activation |
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Jung Kyu Choi, Jae Hee Lee, Seung Tae Lee, Mun Hwan Choi, Seung Pyo Gong, Eun Ju Lee, Jeong Mook Lim* |
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Correspondence:
Jeong Mook Lim,
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| Abstract |
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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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