• 한국수정란이식학회지
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• 제11권3호
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• pp.201-210
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• 1996

This experiment was carried out to evaluate the effect of early mouse embryonic development in vitro by co-culture with bovine and porcine oviductal epithelial cells (BOEC and POEC). The 2-cell embryos were collected from the oviducts of the superovulated and mated cultured in D-PBS /15% FCS at 48 hours after hCG injection. The in vitro developmental rate of blastocyst formation in the embryos were examined under the fllowing treatments; 1) TCM 199 added 15% HCS, 2) Ham's F-10 added 15% HCS, 3) MediCult IVF medium, 4) TCM 199 added 15% HCS + BOEC, 5) TCM 199 added 15% HCS + POEC, 6) Ham's F40 added 15% HCS + BOEC, 7) Ham's F-10 added 15% HCS + POEC,8) MediCult IVF medium + BOEC, 9) MediCult IVF medium + POEC. For a comparative study of in vitro development for 96 hours after hCG injection, were cultured with oviductal epithelial cell and media only. The obtained results were 2-cell embryos developed to the blastocyst stage in TCM 199, Ham's F-10 and MediCult IVF medium at the rates of 84.4,83.2 and 81.6%. respectively. The higher developmental rates(91~97%) of blastocyst formation was appeared when the embryos were co-cultured with a monolayer of bovine or porcine oviductal epithelial cells in TCM 199 or Ham's F-10 and MediCult IVF media. No significant difference in developmental rates was shown between bovine and porcine oviductal epithelial cells but significant difference in co-culture system in comparison between media only system and co-cultures. In conclusions, oviductal epithelial cells, BOEC and POEC, when co-culture with mouse early embryos improved the rates of development, blastocyst and hatching. Therefore, it is suggested that co-culture system using oviductal epithelial cells improve early embryonic developtnent in mouse.

• Asian-Australasian Journal of Animal Sciences
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• 제13권7호
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• pp.894-896
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• 2000

In vitro fertilization can be used for salvaging superior buffalo germplasm which otherwise goes waste after the slaughter of animals. This technology has also increased our basic understanding of growth of germ cells and embryos. The requirement of growing embryos is peculiar and stage specific. In the present study the cleavage and development of buffalo embryos were studied with homologous (buffalo) and heterologous (goat) oviductal cell co-culture systems. The cleavage rate improved significantly (p<0.01) in both homologous and heterologous co-culture as compared to control (55.3, 46.8 and 11.4%). The morula formation using homologous and heterologous oviductal cells also increased significantly as compared to control group (43.6, 21.9 & 1.9%). There was no blastula formation in control group, but addition of oviductal cells either from homologous or heterologous species significantly increased the blastula formation (9.5, 12.5%). The cleavage rate and embryo development was slightly better (non significant) in homologous as compared to heterologous oviductal cell culture. It was concluded that the use of oviductal cell co-culture (homologous and heterologous species) have significantly improved cleavage and development of buffalo embryos in vitro.

• 한국수정란이식학회지
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• 제11권1호
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• pp.7-14
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• 1996

The object of this study was to evaluate the effect of uterine epithelial cells on development of Korean native cattle(KNC) oocytes fertilized in vitro. Qocytes were collected from ovaries of slaughtered Korean Native Cows and matured in TCM199 with granulosa cells supplemented with 10% FBS, 5$\mu$g/ml FSH, 10 JU/ml hCG, and 1$\mu$g/ml estradiol-17$\beta$ for 24 hrs. For co-culture of in vitro development of fertilized ova, oviductal epithelial cells (l$\times$l0˚cells /ml) obtained from slaughtered cow and uterine epithelial cells(1$\times$10˚cells /ml) flushed from the superovulated holstein on Day 7 were incubated in 39$^{\circ}C$, 5% $CO_2$, 95% air. Frozen-thawed KNC sperm was capacitated with BO(Brackett & Oliphant, 1975) medium supplemented with 10mM, 5mM-caffein. Matured oocytes were inseminated for 20 hrs. And then fertilized oocytes were washed with culture medium and transferred to oviductal epithelial cells for in vitro development and three days later a portion of embryos were transferred to uterine epithelial cells. Stastical methods of developmental rates on KNC-IVF oocytes was ANOVA-test. Developmental rates of KNC-IVF oocytes was significant higher(P<0.01) when co-cul-tured with uterine epithelial cells(25.2%) than oviductal epithelial cells. Blatocyst cul-tured for 7 to 9 days were frozen by automatic freezer with 1.4M glycerol-PBS. Survival rates of blastocyst was 40.0%. Fourteen frozen-thawed blastocysts were transferred to five holstein heifers on day 7 after natural estrus. Three recipients were observed twin and one recipient was single by ultra-sound systems on days 45 after embryo transfer.

• 한국가축번식학회지
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• 제19권2호
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• pp.135-140
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• 1995

The study was conducted to investigate on the survival rate of bisected porcine embryos co-cultured in 10% FCS(v/v)＋TCM-199 media containing hormones, oviductal epithelial cells and cumulus cells 0 to 72 hrs after bisection. In vitro survival rate was defined as development rate on in vitro culture or FDA-test. The results are summarized as follows ; 1. The survival rate of bisected porcine embryos cultrued in 10% FCS＋TCM-199 media contaning PMSG, hCG, PMSG＋hCG, hCG＋$\beta$-estradiol 0 to 20 hrs and 20 to 40 hrs were 37.6% and 37.5%, 28.6% and 28.6%, 35.7% and 28.8%, 30.8% and 23.1%, 38.5% and 30.7%, respectively. The survival rate of bisected embryos co-cultured in TCM-199 media containing hormones, oviductal epithelial cells and cumulus cells was significantly higher than that of non co-culture. 2. The survival rate of bisected porcine embryos co-cultured 10% FCS＋TCM-199 media containing oviductal epithelial cells 4 to 5 hrs and 20 to 24 hrs were 42.9% and 38.5%, respectively. 3. The survival rate of bisected porcine embryos co-cultured in 10% FCS＋TCM-199 media containing cumulus cells 4 to 5 hrs and 20 to 24 hrs were 40.0% and 35.7%, respectively.

• 한국가축번식학회지
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• 제18권4호
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• pp.285-297
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• 1995

The mammalian oviduct is a place where ontogeny of an animal begins. Nowadays, however, it is possilbe to manipulate a part of physiological events occurring in the oviduct so that fertilization of gametes and early embryonic development of zygotes could proceed outside oviductal environment. Rabbit zygotes readily develop to blastocysts in a conventional culture condition. Most of the mouse fertilized eggs do so when cultured under a specific environment, e.g., in a medium containing ethylenediamine tetraacetic acid. Similarly, a significant number of zygotes from rat, sheep, pig or cattle can develop to blastocysts if they are cultured in the presence of particular component which appear to be somewhat species-specific. Instead of changing the components of medium, somatic cells including oviductal epithelial cells, have widely been used to improve mammalian embryonic development in vitro. Many investigators have reported that mammalian zygotes, whether fertilized in vivo or in vitro, could develop to blastocysts when they were cultured on a monolayer of various kinds of somatic cells or even in a somatic cell-conditioned medium. While little is known about the nature of embryotrophic factor(s) produced in vitro by somatic cells, the existence fo oviduct-specific protein(s) has consistently been demonstrated in many laboratories. Some of these proteins are reported to be associated with oviductal eggs. However, the physiological role of these proteins has still to be determined. Recently we observed that the perivitelline space of mouse oocytes was fluorescently stained with various fluorochrome-protein conjugates following ovulation into the oviducts or upon their expossure to oviductal extracts. Furthermore, it was also found that cattle or pig oviductal fluid gave similar results when examined using mouse ghost ZP. These observations lead to suggest that mammalian oviduct induces changes of biochemical properties of oocytes. Further studies are needed to clarify the nature of oviductal factor(s) and the physiological meaning of the reaction.

• 한국수정란이식학회지
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• 제10권3호
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• pp.237-242
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• 1995

Porcine follicular oocytes matured in culture were inseminated with frozen-thawed spermatozoa. When the oocytes were inseminated in the medium with oviductal epithelial cell monolayer, the penetration rates higher in those with (4.1, 31.7, 45.1, 54.5 and 69.4%) than without cells (0, 17.1, 34.8, 45.2 and 58.9%) at 4, 8, 12, 16 and 20 h after insemination. The proportions of polyspermy in penetrated oocytes in medium with or without cells increased with time of examine. In another experiment, the penetration rate was higher without (57.6%) than with (19.6~24.1%) preincubation of spermatozoa for 1~4 h in medium. However, when the oocytes were inseminated with spermatozoa preincubated for 1~2 h, the penetration rates significantly higher (P<0.05) in those with (65.6 and 55.9% for 1 and 2 h) than without (24.1 and 20.6% for 1 and 2 h) oviductal epithelial cell monolayer. On the other hand, the proportions of polyspermy decreased with time of spermatozoa preincubation. These results indicate the significant advantages of the spermatozoa preincubation with oviductal epithelial cell monolayer for 1 and 2 h to maintain penetration potential during in vitro fertilization in the porcine.

• 한국가축번식학회지
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• 제19권4호
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• pp.259-264
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• 1996

This study was carried out to investigate on the survival rates and in vitro developmental rates of bisected bovine embryos were by manipulator and micropipette. Bisected embryos were co-cultured in 20% FCS(v/v)＋TCM-199 media containing hormones, oviductal epithelial cells and cumulus cells 0 to 72 hrs after bisection. Survival rate and in vitro fertilization rate were defined as development rate on in vitro culture or FDA-test. The results are summarized as follows ; 1. The in vitro developmental rate of biseciton embryos co-cultured in 20% FCS＋TCM-199 medium containing PMSG, hCG, PMSG＋hCG, hCG＋$\beta$-estradiol 0 to 20 hrs and 20 to 40 hrs were 36.7, 26.7, 33.3, 40.0, and 30.0, and 30.0, 33.3, 30.0, 26.7, and 26.7%, respectively. The survival rate of bisection embryos co-cultured in TCM-199 medium containing hormones was significantly higher than that of non co-culture(25.0%). 2. The in vitro developmental rates of bisection embryos co-cultured in 20% FCS＋TCM-199 medium containing oviductal epithelial cells 4 to 5 hrs and 20 to 24 hrs were 40.0 and 33.3%, respectively. The survival rate of bisection embryos co-cultured in TCM-199 medium containing oviductal epithelial cells was significantly higher than that of non co-culture(25.0%). 3. The in vitro developmental rates of bisection embryos co-cultured in 20% FCS＋TCM-199 medium containing cumulus cells 4 to 5 hrs and 20 to 24 hrs were 43.3 and 36.7%, respectively. The survival rate of bisection embryos co-cultured in TCM-199 medium containing cumulus cells was significantly higher than that of non co-culture (25.0%).

• Applied Microscopy
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• 제10권1_2호
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• pp.41-51
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• 1980

정상(正常) 성주기(性週期)를 나타내는 백서(白鼠)의 난관(卵管)에서 각(各) 영역(領域)에 분포(分布)하는 상피세포(上皮細胞)를 주사전자현미경(走査電子顯微鏡)으로 관찰(觀察)하여 다음과 같은 결과(結果)를 얻었다. 즉, 백서난관상피세포(白鼠卵管上皮細胞)는 섬모세포와 무(無)섬모세포로 구성되며, 무(無)섬모세포는 4가지 세포형(細胞型)으로 구별(區別)할 수 있고 또한 이들 각형(各型)의 세포(細胞)는 성주기시(性週期時) 특정영역(特定領域)에 분포(分布)하였다.

• 한국가축번식학회지
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• 제20권2호
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• pp.179-190
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• 1996

Sexing and developing from splitted embryos which were fertilized in vitro implicate a possibility of production of the superior and sex controlled individuals. This study was carried out to investigate the production of transferable late blastocysts from in vitro fertilized embryos and to analyze sex by chromosome analysis from same embryos. In results, the ratio of cleavage and fertility of bovine follicular oocytes matured in vitro was 90% in co-cultured with granulosa cells. The competence of embryonic development from in vitro matured and fertilized bovine oocytes was 38% in co-cultured with bovine oviductal epithelial cells. To produce a lot of transferable embryos, therefore, the best conditon of culture system was co-cultured with granulosa cells for immature bovine oocytes and then co-cultured with bovine oviductal eptithelial cells for matured and fertilized oocytes. In chromosome analysis, 93% of in vitro fertilized embryos were very important aspect in chromosome preparation from bovine embryos such as duration of colcemid treatment, weakening of zona pellucida, methods of hypotonic treatment and fixation treatment.

• Asian-Australasian Journal of Animal Sciences
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• 제22권2호
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• pp.194-200
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• 2009

The aim of this study was to investigate the effects of donor cell passage, size and type on the development of nuclear transfer embryos. Porcine cumulus cells, fetal fibroblasts and oviductal epithelial cells from 1-2, 3-6 and 7-10 passages were used for the nuclear transfer. In the oocytes with the cumulus donor cells, fusion and cleavage rates of oocytes and cell numbers per blastocyst among the three different passage groups did not show any differences, but the rates of blastocyst formation from 1-2 and 3-6 passage groups were higher than those from 7-10 passage group. The rates of fusion, cleavage and blastocyst formation, and the cell numbers per blastocyst were higher in the embryos with the sizes of <20 and 20 ${\mu}m$ cumulus donor cells compared to the >20 ${\mu}m$ cumulus donor cell. In the oocytes with the fetal fibroblast donor cells, the rate of blastocyst formation from the 3-6 passage group was higher than from 1-2 and 7-10 passage groups. The embryos with the size of 20 $\mu{m}$ fetal fibroblast donor cell showed higher rate of blastocyst formation compared to those with <20 and >20 ${\mu}m$ donor cells. In the oocytes with the oviductal epithelial cells, the rates of blastocyst formation from 1-2 and 3-6 passage groups were higher compared to those from 7-10 passage group. The embryos with the sizes of <20 and 20 ${\mu}m$ oviductal epithelial donor cells had a higher rate of blastocyst formation compared to those with >20 ${\mu}m$ donor cell. Fusion and cleavage rates of oocytes, and cell numbers per blastocyst among the three different donor cell types from the 3-6 passage did not show any differences. However, the rate of blastocyst formation of somatic cell nuclear transfer (SCNT) embryos with the fetal fibroblast donor cell was higher than that of blastocyst formation of SCNT embryos with the cumulus and oviductal epithelial donor cells.