• Journal of Embryo Transfer
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• v.19 no.3
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• pp.229-237
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• 2004

The main objective of this study was to examine the effects of serum and gonadotropins supplement during in vitro maturation(IVM) of bovine oocytes on nuclear maturation and embryo development, and we also examine the cell number. 1 . The first polar body(PB) extrusion rates of Korean native cow(KNC) oocytes matured in medium with FBS or gonadotropins were similar among treatment groups. The development rate to the blastocyst stage was significantly higher in the group of both supplement FBS and gonadotropins(26.0％) than in the group of non-supplement(9.9％) and gonadotropins (12.0％). The numbers of inner cell mass (ICM) and trophectoderm (TE) cells and total cell numbers of blastocysts were highest in the group of both supplement FBS and gonadotropins, and the number of ICM cells was increased by FBS supplementation (p<0.05). 2. The PB extrusion rates of KNC oocytes matured in medium with FBS in the different duration of IVM was significantly higher in the 0-18hr(63.1％) and in the 9-18hr(63.4％) group than in the 0-9hr.(37.4％) group (p<0.05). The embryo development rates did not differ among treatment groups. The numbers of TE cells and total cell numbers of blastocysts were similar among treatment groups, but the number of ICM cells of the 0-18h. group were significantly higher than the other treatment groups (p<0.05). The results indicate that although TCM199 alone can support bovine oocyte maturation and development to the blastocyst stage, a high quality of blastocysts can be produced from oocytes matured in medium containing serum and gonadotropins.

• Journal of Embryo Transfer
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• v.21 no.2
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• pp.157-162
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• 2006

It is well known that unidentified factors in sera, hormones and growth factors promote the proliferation of granulosa cells and nuclear maturation of bovine COCs (cumulus oocytes complexes) in vitro. Attempts had been developed the simple composition of culture media and similar system to in vivo conditions has been applied. In the present study, we investigated the effect of FGF (fibroblast growth factor) on in vitro maturation and in vitro development of Hanwoo COCs. When the COCs were matured in HPM 199 (Inst. of Functional peptide, Japan) containing 0.1, 1 and 10 ng/ml FGF for 24 hr, maturation rates to metaphase II ($70.0{\sim}75.0%$) were significantly higher (p<0.05) than that of control group (0 ng/ml FGF, 37.5%). When matured COCs with FGF were cultured in maturation medium after in vitro fertilization, developmental rates to blastocysts were 9.5, 0 and 2.9%, respectively, compared to 25.0% of the control group (p<0.05). When the matured COCs with FGF were cultured in HPM 199 (IFP971, Inst. of Functional peptide, Japan) containing 10% FBS, 0.8% BSA or 0.1% PVA (polyvinyl alcohol), the blastocyst formation rates were 12.4, 12.8 and 8.5%, respectively, while the rates of matured COCs with FGF and cultured with IVMD and IVD (Inst. of Functional peptide, Japan) without serum were 38.4% and 34.8%, respectively (p<0.05). These results suggested that FGF is available for in vitro maturation of bovine COCs and is not suitable for in vitro development, but further investigation would be need for finding the synergistic autocrine/paracrine fashion of other growth factors in early bovine embryo development.

• Korean Journal of Animal Reproduction
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• v.24 no.1
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• pp.97-108
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• 2000

The objectives of this study were performed to increase the efficiency of the culture conditions of embryos produced in vitro, and to assess the developmental potential after transfer of those embryos into recipients. The mean number of folliclular oocytes recovered from an ovary was 10.7. The rates of maturation and fertilization in Grade I oocytes were significantly (P＜0.05) higher than Graden and III. Developmental rate into blastocyst in the culture group of TCM-199 with BOEC were significantly higher (P＜0.05) than the groups of TCM-199 and conditioned medium (24.7% vs. 12.4% and 18.2%). The survivability of post-thawed blastocysts equilibrated for 3 min in EFS solution was significantly (P＜0.05) lower than l0 for 1 and 2 min (32.1% vs. 82.9% and 73.3%). Significantly higher (P＜0.05) survival rate in blastocysts was seen after freezing than in morulae stage embryos. Out of all 105 recipients, 49 (46.7%) were confirmed in pregnant. On pregnancy of cattle, 48 calves were born from 40 recipients. The ratio of twin and single calves was 30.5% (32/40 and 7.6% (8/40), respectively. However, the others composed of abnormal, as judging as 6 (12.2%) for abortion and 3 (6.1%) for stillbirth during the pregnant period.

• Journal of Embryo Transfer
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• v.16 no.3
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• pp.193-201
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• 2001

This study was designed to examine the factors affecting in fertilization and development of embryos in vitro, and to examine whether zone drilling by laser irradiation can improve the hatching rate of IVF embryos from DNA marker-proved Hanwoo. DNA markers related to marbling score were identified using DNA fingerprinting with Ml3 probe and restriction enzyme Hae III. Oocytes were aspirated from immature ovarian follicles using a combined method of rectal ovarian-palpation and transvaginal ultrasound-guidance(6.5MHz) under local anesthesia. The aspirated oocytes were washed twice with fresh D-PBS containing 5% FBS and were rewashed 4 to 5 times with TCM-199 containing 5% FBS. A morphological grade of I to IV was assigned to each oocyte. Data were analyzed using the GLM procedure of SAS. Sperm separation methods did not have any significant effect on cleavage or developmental abilities of IVF embryos. Significantly(P<0.05) higher cleavage rate was observed in embryos from GI(60.0%, 3/5), GII(69.2%, 18/26) and GIII(62.1%, 59/95) compared to embryos from GIV oocytes(36.2%, 25/69). And the developmental rate to blastocyst stage was higher(P<0.05) in embryos from GI(33.3%, 1/3) and GII oocytes(38.9%, 7/18) than those from GIII(16.9%,10/59) and GIV oocytes(4.0%, 1/25). There was no significant difference in development of IVF embryos to blastocyst by media for in vitro culture. Proportion of hatched blastocyst was significantly(P<0.05) higher in embryos received zona drilling by laser than those of non-drilled.

• Development and Reproduction
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• v.8 no.2
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• pp.119-122
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• 2004

It is well known that unidentified factors in sera, hormones and growth factors promote the proliferation of granulosa cells and nuclear maturation of bovine COCs in vitro. Attemps had been developed the simple composition of culture media and similar system to in vivo conditions has been applied. In the present study, we investigated the effect of TGF-${\beta}$ on in vitro maturation and in vitro development of Hanwoo COCs. When the COCs were matured in TCM 199 containing 0.1, 1 or 10 ng/ml TGF-${\beta}$ for 24 hrs, metaphaseⅡ of COCs were obtained 95.8%, 100% of matured COCs, respectively and there were no differences among the concentrations of TGF-${\beta}$. Matured COCs with TGF-${\beta}$ cultured in maturation medium after in vitro fertilization, developmental rate to blastocyst were 0~0.8%. Matured COCs with TGF-${\beta}$ were cultured in TCM 199+10% FBS, 0.8% BSA, 0.1% PVA, blastocyst formation were showed in 12.4%, 12.8%, 8.5% of those and cultured in IVMD or IVD without serum were 38.4%, 34.8%, respectively. There were significant differences among the media (P<0.05). TGF-${\beta}$ is available for i vitro maturation of bovine COCs, but further investigation would be need for finding the synergistic autocrine/paracrine fashion of other growth factors in early bovine development.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2005.10a
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• pp.124-124
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• 2005

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2005.10a
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• pp.124-124
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• 2005

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2004.10a
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• pp.72-72
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• 2004

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2004.10a
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• pp.97-97
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• 2004

• Korean Journal of Animal Reproduction
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• v.24 no.1
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• pp.83-88
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• 2000

This study was conducted to determine the effect of repeated ultrasound-guided transvaginal retrieval of oocytes recovery, estrous cycle and ovarian adhesion in Korean native, Hanwoo heifers. Heifers were at unknown stages of the estrous cycle at the start of experiments in which all follicles $\geq$6mm in diameter were ablated. The results obtained in this study were as follows; Follicle developing number and oocytes collected number were no effected to repeated OPU to nine session, 4 e.a range oocytes collected to repeated OPU session. Oocytes were observated follicles were 8.7$\pm$4.2 e.a, collected oocytes were 4.1$\pm$3.4 e.a to two times collected per week and observated follicles was 10.2$\pm$6.1 e.a, collected oocytes were 4.3$\pm$2.9 e.a to one times collected per week, but no difference significantly(P＜0.05). Ovaries adhesive percentage to repeated OPU was eight ovaries adhisived(20%) of forty ovaries, three ovaries(7.5%) to 1~3 times oocytes collected, four ovaries(l0%) to 4~6 times, one ovaries(2.5%) to 7~9 times oocytes collected session. To repeated OPU effection, ovaries adhisive heifers were long estrous cycle(＞25 day) to 7 heads(87.5%) of 8 head, non-adhesive ovaries heifers were 5 heads(41.7%) were long estrous cycle to repeated OPU 12 heads. Although, now unknown about the dynamics of follicles wave and about functional changes to repeated OPU ovaries, more question about ovaries adhesive cause remain.