Influence of Ovulation Induction Medicine on the Nuclear Maturation of Mouse Immature Oocytes and Development of Mouse 2-cell Embryo in Various Culture Media> / 대한불임학회지

Purpose of the present study was to find the optimal ovulation induction medicine for the maturation and development of immature oocytes and culture media for 2-cell embryos in the mouse model. ICR female mouse aged 6 to 8 weeks, were stimulated with 5 IU PMSG injection. At 47 to 50 hour post-PMSG injection, ovaries were dissected out and oocytes-cumulus complexes were punctured. The oocyte-cumulus complexes were cultured in media containing various ovulation induction medicine, CC, HMG and Metrodin for 18 hours. Female ICR mice were stimulated with 5 IU PMSG and 48 hours later were injected 5 IU of hCG, then female and male mice were mated. At 48 hour post-hCG injection, oviducts were dissected out and 2-cell embryos were flushed. The 2-cell embryos were cultured in various media, Ham's F-10 media of milli-Q water (3degrees), Ham's F-10 media of HPLC (high performance liquid chromatography, Baxter) water, Medicult media, HTF (human tubal fluid) media for 96hours. The results were as follows. 1. When the oocytes-cumulus complexes were cultured in 10(-9)microgram/ml~ 10(-8)microgram/ml of CC, those were suppressed in meiotic maturation (28.2~ 33.7%). Whereas the oocytes-cumulus complexes were cultured in 10(-7)microgram/ml~10(-4)microgram/ml, these were not effected in meiotic maturation (54.5~72.7%). 2. When the oocytes-cumulus complexes were cultured in 10(-4)microgram/ml~ 10(-1)microgram/ml of Metrodin, those were suppressed in meiotic maturation (35.7~ 41.5%). Meanwhile the oocytes-cumulus complexes were cultured in 10(-7)microgram/ml~10(-5)microgram/ml, those were not effected in meiotic maturation (54.2~ 70.3%). 3. When the oocytes-cumulus complexes were cultured in 10(-5)microgram/ml~ 10(-4)microgram/ml of HMG, those were suppressed in meiotic maturation (48.2~ 50.4%). As being cultured in 10(-7)microgram/ml~10(-6)microgram/ml, increased in meiotic maturation (75.8~80.7%). 4. When the 2-cell embryos were cultured in Ham's F-10 media of milli-Q wats. ( 3degrees), Ham's F-10 media of HPLC (high performance liquid chromatograpy, Banter) water, Medicult media, HTF (human tubal fluid) media, developmental rates to blastocyst and hatching for 96 hour were 50.0%, 45.2%, 71.5% and 95.6%, respectively.

The Evaluation of Various Conditions in the Cryopreservation of Mouse Embryos - Rapid and Slow Method of Cryopreservation, Culture Media and Cell Stages / 대한불임학회지

Cryopreservation is able to store the surplus pre-embryos for freezing and furthermore thawing and transfer in a subsequent cycle. Cryopreserving cells which are maintaining their viability are the very complex process. This study has been carried out in order to find the effects of cryopreservation steps, freezing media and embryonic stages on the rates of viability and development of cryopreserved mouse embryos. Female ICR mice (6~8 weeks old) were induced to superovulate by sequential intraperitoneal injection of 5 IU PMSG and 5 IU hGC 48h apart. Mouse embryos were collected according to its developmental stage after the injection of hCG. Embryos were cryopreserved not only by cryoprotectant step (1 step~ 4 step) but also in a variety of media (HTF, IVF medium, D-PBS) and cell stage. The results were as follows: There is no clear advantage in these freezing media of rapid method, but 4 cell and 8 cell of slow method (2, 3, 4 step) have advantage in D-PBS. The development of embryos according to cell stage become greater in 8 cell stage. In the treatment steps of cryopreservation, the development of embryo to blastocyst was similar among rapid method, but the development of 4 cell and 8 cell embryos to blastocyst according to slow method was better than rapid method.