• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.20-20
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• 2001

It has been reported that cloning cattle is inefficient. One of the problems was placental abnormality, finally resulting in fetal mortality after transfer of nuclear transfer (NT) bovine embryos. This study was focused on the allocations of embryonic cells to the inner cell mass (ICM) or to the trophectoderm(TE) in NT bovine blastocysts. Somatic cells were derived from a Day 45 fetus of gestation, individually transferred into enucleated oocytes and developed to the blastocyst stage in vitro. Differential staining was used to assess the qualify of blastocysts derived from NT, IVF and in vivo. Development rate of NT embryos to blastocysts (25.0%, 41/164) was similar to that of IVF embryos (28.7%, 49/171). The total cell number of NT blastocysts (101.3$\pm$45.9) was not different compared with that of IVF embryos (107.9$\pm$34.2, P>0.05), but was lower than in vivo embryos (122.5$\pm$21.6, P<0.05). Ratio of ICM/total cells was higher in NT embryos (51.6$\pm$ 18.6%) than in IVF and in vivo embryos (42.3$\pm$ 15.3% and 34.9$\pm$8.9%, respectively) (P<0.05). Most IVF (56.8%, 25/44) and in vivo blastocysts(80.8%, 21/26) was distributed in the proportion of ICM/total cells ranging from 20 to 40% group. However, most NT blastocysts was biased in the 40-60%(34.1%, 15/44) and >60% (31.8%, 14/44) groups. Our findings suggest that placental abnormalities or early fetal losses in the present cloning system may be due to aberrant allocation of NT embryos to the ICM cells.

• Korean Journal of Animal Reproduction
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• v.6 no.1
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• pp.19-30
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• 1982

1. The cows slaughtered at age of 3, 4, 6, 7, 8, and 9 years old were 1.5, 1.5, 15.0, 62.5 and 4.4% respectively. 2. The cows slaughtered at 351-450kg and more than 500kg were 60 and 28% respectively. 3. Best, very good, good and bad cows in nutritional condition were 1.6, 25.8, 62.9, and 9.7% respectively. Among the six cows which were bad nutrition, the two were with severe endometritis, the three were normal in genital function and one was on 70 days of pregnancy. 4. Holstein cows(55.2%) showed higher reproductive failure than the Korean cows(33.3%). 5. The slaughted ratio of the Korean cattle and Holstein cows was 36 and 64% respectively. 6. Pregnant cows were about 16% among the slaughtered one. 7. Reproductive failures were composed of 46% in uterus, 32% in ovaries, 8% in udder, 6% in oviduct, 4% in cervix of uterine, 2% in vagina and 2% inmummified fetus. 8. Forty six percentages of uterine diseases were as follows; horn, 13%, body of uterus, 32% and ovary diseases were 32%, that is, 12% of ovary atrophy, 8% of ovarycyst and 6% of lutealcyst. 9. The cows of reproductive failures were commonly infected with 1.6 kinds of diseases. 10. According to classification, six type of ovaries were as follows; normal, 58%, ovary-cyst, 11%, luteum cyst, 4%, coexistence of follicles and corpus luteum, 16%, weak function of ovaries, 10% and ovarian atrophy, 1%. 11. Major axis, minor axis and thickness of right ovary were larger than those of left one both in Korean cattle and Holstein cows. Holstein cow had generally larger size of ovary than these of the Korean cattle.. 12. The left and right oviducts showed no difference in length, but Holstein had longer oviduct than Korean cow. 13. There was no difference in the length of uterine horn between right and left in the Korean cows, but the right was longer than the left in Holstein cows. 14. Holstein had longer horn and body of uterine than the Korean cows. 15. The weight of right ovary was heavier than that of left in both breeds, but there was no differences in weight of left ovary between two breeds and right ovary of Holstein breed was heavier than that of the Korean cow. 16. The weight of right oviduct and uterine born was heavier than that of the left, and Holstein had heavier oviducts and uterine horns than the Korean cows. 17. Holstein had heavier uterine body and cervix of uterine than the Korean cows. 18. The length of reproductive systems of Korean cow is as follows; Major and minor diameter and thickness ofovary are 3.6${\pm}$0.7, 2.3${\pm}$0.4 and 1.6${\pm}$1.4 cm in left and 3.7${\pm}$0.6, 2.5${\pm}$0.5 and 1.8${\pm}$0.5 cm in right. Oviduct is 28.4${\pm}$3.1 cm in left and 27.8${\pm}$3.3 cm in right. Uterine horn is 27.4${\pm}$4.5 cm in left and 27.7${\pm}$4.9 cm in right. Uterine body and cervix are 3.4${\pm}$1.1 and 6.5${\pm}$1.7 cm. 19. The length of female reproductive systems ofHolstein cow is as follows; Major and minor diameter and thickness of ovary are 3.9${\pm}$1.3, 2.3${\pm}$0.5, and 1.5${\pm}$0.6 cm in left and 4.0${\pm}$0.8, 2.8${\pm}$0.6 and 1.8${\pm}$0.6 cm in right. Oviduct is 29.4${\pm}$4.2 cm in left and 29.3${\pm}$4.1 cm in right. Uterine horn is 30.2${\pm}$7.4 cm in left and 32.6${\pm}$8.4 cm in right. Uterine body and cervix are 4.5${\pm}$2.5 and 7.8${\pm}$2.9 cm. 20. The weight of reproductive systems of Korean cow is as follows; Ovary is 8.4${\pm}$4.1 g in left and 9.3${\pm}$3.6g in right. Oviduct is 1.5${\pm}$0.5 g in left and 1.6${\pm}$0.5 g in right. Uterine horn is 109${\pm}$27 g left and 118${\pm}$32 g in right. Uterine body and cervix are 30.4${\pm}$14.1 and 76.7${\pm}$38.4g. 21. The weight of reproductive systems of Holstein cow is as follows; Ovary is 8.2${\pm}$3.1 g in left and 12.5${\pm}$5.6 g in right. Oviduct is 1.7${\pm}$0.6 g in left and 1.9${\pm}$0.9 g in right. Uterine horn is 199${\pm}$14.2 g in left and 221${\pm}$111.2g in right. Uterine body and cervix are 58.2${\pm}$46.5 and 126.7${\pm}$103.3 g.

• Journal of Embryo Transfer
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• v.6 no.2
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• pp.1-17
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• 1991

It is widely recognized that the embryonic or fetal loss after breeding is common in the cattle and that it is an important factor affecting reproductive efficiency. The causes of this loss have been subject of extensive researches and the results indicate that the embryonic mortality may he primary factor responsible for low pregnancy rates in non-embryo transfer bovine populations as well as embryo transfer programs. However, it's causes are still not clearly understood. The embryonic mortality or pregnancy rate has been influenced by various embryonic and maternal effects related to genetic and environmental factors. The timing and extent of embryonic mortality vanes greatly according to authors and estimating methods, because it is difficult to make direct measurements. The major important factors that may influence the embryonic losses or pregnancy rates after embryo transfer can be summeirized. 1.When an embryo is transferred to unmated recipients, the contralateral transfer to corpus luteum results in a lower survival rate than ipsilateral deposition. When the embryos are transferred for the production of twin calves, their survivals and twin pregnancies have quite inconsistent according to the transfer methods either to the unmated-synchronized or already mated recipients and more works are needed to accurrately clarify the previous results. 2.Although embryos can be cultured in vitro some hours without the great declines in pregnancy rates, the rates differ markedly among culture times and media but may be improved by co-transfer systems. 3.Embryo developmental stages and quality grades clearly affect the survival rate following freezing and the pregnancy rate after transfer and the selection of embryos without chromosome abnormalities and of high fertile semen may also be considered to increase the pregnancy rates. 4.Many researches have attempted to relate the plasma progesterone levels to pregnancy rates and others have done either direct progesterone supplementation or luteal stimulation by hCG treatment in order to increase the pregnancy rates. However, these effects on pregnancy rates are inconsistent and also contradictory. 5.The asynchrony between donors or embryos and recipients may he a major cause of embryo death and low pregnancy rate and the sensitivity to uterine asynchyony differs in according to the quality and stages of embryos. 6.The extremes of poor or over nutrition during early pregnancy in the recipients are detrimental to the survival of embryos and the good body condition is required to prevent a reduejion of pregnancy rates. The uterine pathogens in embryonic mortality or fertility have been questioned but the infection of C.pyogenes and Campylobacter fetus is still important pathogens. 7.The heat stress during early pregnancy may reduce conceptus weight and possibly increase the embryonic mortality.

• Reproductive and Developmental Biology
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• v.30 no.1
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• pp.53-58
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• 2006

The study was conducted to evaluate the effects of culture period and fusion method on the development of somatic cell nuclear transfer (SCNT) embryos reconstituted with Korean bovine fetal fibroblast cells (KbFF) and Korean bovine adult ear skin fibroblast cells (KbESF). KbFF were isolated from a day 51 Korean cattle (Hanwoo) fetus, and KbESF were isolated from a 28 month old Hanwoo calf. The cells were cultured up to 15 weeks (passage 15) in vitro for SCNT. Chamber and electrode needles were used for comparing fusion of reconstituted eggs. The doubling times of KbFF and KbESF were 17.3 hr and 24.3 hr, respectively. The fusion and cleavage rates were significantly higher in needle group (76.1 and 81.2% respectively, P<0.05) than those in chamber group. However, the blastocyst development rate was not different between both groups. Fusion and cleavage rates of NT eggs reconstituted with KbESF did not affected by passage number, however, blastocyst rates were lower in passage $1{\sim}4$ group (21.3%) than passage $5{\sim}8$ (39.4%) and $13{\sim}15$ groups (40.4%, P<0.05). Whereas, fusion rate was lower in passage $1{\sim}4$ group (61.5%) than those of passage $5{\sim}8$(75.0%) and $13{\sim}15$ (76.8%) groups, but cleavage and blastocyst rates were similar regardless of passage number in the KbFF. The results suggest that fusion method can affect the development of SCNT embryos, whereas the long term culture up to 15 passages may not affect the development of SCNT embryos.

• Journal of Embryo Transfer
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• v.21 no.1
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• pp.13-20
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• 2006

The purpose of this study was to develope the transgenic cattle expressing hFSH into the urine using the nuclear transfer. To produce the interest gene in urine, the specific vector was ligated with hFSH gene undo. maUII promoter. The fetal fibroblast cells (KbFF) were isolated from a 45-day male fetus. The hFSH gene was co-transfected with pcDNA3 (neo) vector to KbFF cells by electroporation. The gene-transfected cells were cultured with G-418 selection medium for 2 weeks. Selected colonies were confirmed by PCR. For nuclear transfer, enucleated bovine oocytes were transferred with hFSH transfected or nontransfected fetal fibroblasts. The cleavage and blastocyst formation rates were significantly lower (p<0.05) in cloned embryos transfected with hFSH gene (68.7% and 15.7%) than in those non-transfected (67.6% and 24.5 %), respectively. Apoptosis analysis showed no difference between hFSH transfected and non-transfected blastocysts (p>0.05). The blastocysts were transfected to 77 (control 24, hFSH 53) recipient cows. Two calves were born (1.9%) following transfer with NT embryos transfected with hFSH gene, but they were confirmed not to be transgenic calves. This result shows that the hFSH colonies were mixed with transfected and non transfected cells. Further research will be needed for selection and establishment of gene transfected cells.