Reproductive medicine and biology · 2020
Usefulness of modified Medium RD as a chemically defined medium for in vitro maturation of bovine oocytes.
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Momozawa Kenji
Аннотация
PURPOSE: In the present study, I evaluated the usefulness of Medium RD, with mixed RPMI1640 and Dulbecco's modified Eagle's medium (1:1, v/v), as a chemically defined medium for in vitro maturation (IVM) of bovine oocytes.
METHODS: In vitro maturation was performed in 10 mmol/L HEPES-buffered TCM199 (mTCM199), 10 mmol/L HEPES-buffered Medium RD (mRD), and mTCM199 supplemented with fetal bovine serum fraction (mTCM199 + FBS fraction) that served as control. Cumulus-oocyte complexes were matured for 24 hours in three different media supplemented with follicle-stimulating hormone, estradiol-17β, and polyvinylpyrrolidone. Nuclear maturation of oocytes, their developmental competence into blastocysts after in vitro fertilization (IVF) and mitochondrial distribution in oocytes were investigated.
RESULTS: There was no difference in the ratio of matured oocytes regardless of IVM media. The percentage of morula stage was higher in mRD than in mTCM199 group (P < .05) at 120-144 hours after IVF, although the blastocyst rates between groups were not significantly different at 168-216 hours. IVM in mRD increased the percentage of oocytes with diffused mitochondrial distribution compared with the immature and mTCM199 and had similar percentage of oocytes in mTCM199 + FBS fraction.
CONCLUSIONS: Medium RD would be useful as a chemically defined medium for IVM of bovine oocytes.
METHODS: In vitro maturation was performed in 10 mmol/L HEPES-buffered TCM199 (mTCM199), 10 mmol/L HEPES-buffered Medium RD (mRD), and mTCM199 supplemented with fetal bovine serum fraction (mTCM199 + FBS fraction) that served as control. Cumulus-oocyte complexes were matured for 24 hours in three different media supplemented with follicle-stimulating hormone, estradiol-17β, and polyvinylpyrrolidone. Nuclear maturation of oocytes, their developmental competence into blastocysts after in vitro fertilization (IVF) and mitochondrial distribution in oocytes were investigated.
RESULTS: There was no difference in the ratio of matured oocytes regardless of IVM media. The percentage of morula stage was higher in mRD than in mTCM199 group (P < .05) at 120-144 hours after IVF, although the blastocyst rates between groups were not significantly different at 168-216 hours. IVM in mRD increased the percentage of oocytes with diffused mitochondrial distribution compared with the immature and mTCM199 and had similar percentage of oocytes in mTCM199 + FBS fraction.
CONCLUSIONS: Medium RD would be useful as a chemically defined medium for IVM of bovine oocytes.