• Journal of Yeungnam Medical Science
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• 제12권1호
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• pp.124-134
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• 1995

Ham's F-10 기본 배양액과 이 배양액에 여러가지 농도의 EDTA와 태아제대혈청을 단독 또는 같이 혼합한 배양액을 얻어 각각의 배양액에서 5-10주된 백서를 이용하여 2 세포기 배아의 단계적 분할정도를 96시간 동안 관찰하였다. Ham's F-10 기본 배양액에 비해 태아제대혈청이나 EDTA $50{\mu}M$에서 $100{\mu}M$을 단독 또는 함께 첨가한 배양액에서 2 세포기의 배아의 상실배기 난할률이 매우 높았으며 (p<0.05), 포배기 난할률은 태아제대혈청과 EDTA $50{\mu}M$에서 $100{\mu}M$을 첨가한 배양액에서 유의하게 높았다(p<0.05). 체외수정실에서 흔히 사용되는 태아제대혈청을 첨가한 배양액과 비교시 태아제대혈청과 EDTA $100{\mu}M$을 첨가한 배양액에서 상실기 난할률이 유의하게 높았다(p<0.05). 태아제대혈청과 여러 농도의 EDTA를 첨가한 배양액과 EDTA만 첨가한 배양액의 비교에서 태아제대혈청과 EDTA $50{\mu}M$ 및 $100{\mu}M$을 첨가한 배양액에서 EDTA $200{\mu}M$만 첨가한 배양액에 비해 상실기 난할률이 매우 유의하게 높았고(p<0.05), 포배기 난할률은 태아제대혈청과 EDTA $100{\mu}M$을 첨가한 배양액이 EDTA $200{\mu}M$을 단독 또는 태아제대혈청과 함께 넣은 배양액에 비해 유의하게 높았다(p<0.05). 결론적으로 Ham's F-10 기본 배양액에 태아제대혈청과 적정 농도의 EDTA $50{\mu}M$ 및 $100{\mu}M$을 첨가한 배양액에서 초기 수정란의 분할이 우수함을 알 수 있고, 이는 수정란의 배양시 적절한 배양액을 선택할 수 있는 기초 자료로서 유용할 것으로 사료된다.

• Clinical and Experimental Reproductive Medicine
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• 제23권1호
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• pp.95-102
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• 1996

This study was performed to investigate the effect of human follicular fluid (HFF) on development of mouse embryos, for evaluating the suitability of HFF as a substitutive material of human fetal cord serum in ART program. The various concentrations of HFF were added into the culture medium and the effects of HFF concentrations were examined to identify the optimal concentration of HFF for embryo development. The potency of HFF in improving embryo development was compared to that of other protein supplement. Collected HFFs were classified with the maturity of the containing oocytes; mature, immature, atretic, and then the effects of the classified HFFs on embryo development were examined. Also, HFF was separated into the low (<30,000 Da) and high (>30,000 Da) molecular weight fractions and the effects of the fractions on embryo development were investigated. The highest development rate was found in culture medium supplemented with 20% HFF, bnt this rate was reversely reduced at the concentrations of HFF higher than 20%. The development rates to the blastocyst, hatching blastocyst, attachment and outgrowth cultured in mature HFF was significantly higher than those in immature and atretic HFF, and mean cell number in blastocyst was higher in mature HFF than in immature and atretic HFF. The development rates of mouse embryos according to protein sources were significantly higher in HFF than in fetal cord serum (FCS), maternal serum (MS) and bovine serum albumin (BSA), and mean cell number in blastocyst cultured in HFF was higher than that in FCS, MS and BSA. The development rates of embryo and mean cell number in blastocyst cultured in high molecular weight fraction of HFF were higher than those in low molecular weight fraction, but the results of high molecular weight fraction were lower than those of whole HFF. Therefore, these results indicated that human mature follicular fluid was useful for improving the development of mouse embryos, which suggests a possibility that HFF also may be used efficiently for improving the culture condition in human ART program as a protein supplement.

• 대한의생명과학회지
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• 제21권1호
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• pp.15-22
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• 2015

Dexamethasone, a synthetic glucocorticoid (GC), is clinically administered to woman at risk for premature labor to induce fetal lung maturation. However, exposure to repeated or excess GCs leads to intrauterine growth restriction (IUGR) and subsequently increases risk of psychiatric and cardio-metabolic diseases in later life through fetal programming mechanisms. GCs are key mediators of stress responses, therefore, maternal nutrient restriction or psychological stress during pregnancy also causes negative impacts on birth and neurodevelopment outcome of fetuses, and other congenital defects, such as craniofacial and skeletal abnormalities. In this study, to examine the effect of prenatal stress on fetal skeletal development, dexamethasone (1 mg/kg [DEX1] or 10 mg/kg [DEX10] maternal body weight per day) was administered intraperitoneally at gestational day 7.5~9.5 and the skeletons were prepared from embryos at day 18.5. Seven out of eighteen (39%) embryos treated with DEX10 showed axial skeletal abnormalities in either the T13 or L1 vertebrae. In addition, examination of the sternum revealed that xiphoid process, the protrusive triangular part of the lower end of the sternum, was bent more outward or inward in DEX group embryos. In conclusion, our findings suggest a possible link to the understanding of the effect of uterine environment to the fetal skeletal features.

• 한국임상수의학회지
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• 제16권1호
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• pp.128-137
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• 1999

Plasma progesterone (P$_4$) concentrations were measured for confirming the estrus observation and for the early pregnancy diagnosis in 130 cows of small farmers. Ultrasonographic examinations were performed from day 30 after artificial insemination to establish the characteristic ultrasonographic appearances of gestational structures in each pregnant stages. Of the 130 cows inseminated, 111 cows (85.4%) were an ovulatory estrus, 12 cows (9.2%) were an unovulatory estrus, and 7 cows (5.4%) were the error of estrus detection, respectively. The accuracy for early pregnancy diagnosis in 111 ovulatory estrus cows achieved when the discriminatory concentration at day 21 after artificial insemination was placed at 3.0 ng-/ml in plasma, was 86.7 % for positive diagnosis and 100% for negative diagnosis, respectively. Pregnancy diagnosis by ultrasonography were performed to evaluate gestational structures from day 30 after artificial insemination in 83 cows. Pregnant cows were 72 of 83 cows. The characteristic ultrasonography of gestational structures in each gestational stages was as follows. The embryo proper was observed within anechoic fetal fluid between 28 and 40 days after insemination, and amnion and embryonic heartbeat was also detected in this period. Between days 41 and 50, embryo proper was detected as an discriminated from head and body, and forelimb buds and hindlimb buds were also observed in this period. Between days 51 and 60, an embryo proper was clearly discriminated from head and body, and fetal movement, forelimb buds and hindlimb buds were observed in this period. Between days 61 and 70, fetus was completely developed, and fetal skeleton, organs and cotyledon were observed. After day 71, each organs of fetus were rapidly developed and a fetus was partially observed in screen because fetus was too big and larger, These results indicate that plasma P$_4$ determination at days 0,6 and 21 after artificial insemination can be utilized for confirming the estrus observation and for early pregnancy diagnosis. Also, ultrasonography was reliable method for early pregnancy diagnosis at day 30 after artificial insemination.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 2003년도 Annual Meeting of KSAP : International Symposium on Pharmaceutical and Biomedical Sciences on Obesity
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• pp.87-87
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• 2003

Taurine (2-aminoethanesulfonic acid, $^{+}NH_3CH_2CH_2{SO_3}^{-}$) is endogenous $\beta$-amino acid which is essential in fetal nutrition and development and is present in abundant quantities in several tissues of fetus. In utero, taurine deficiency causes abnormal development and abnormal function of brain, retina, kidney and myocardium. Thus, transfer of taurine into fetus is important during embryonic development. Taurine transporter (TauT) has 12 hydrophobic membrane -spanning domains, which is typical of the $Na^{+}$- and $Cl^{-}$-dependent transporter gene family. Among the various biosynthetic enzymes of taurine, cysteine sulfinic acid decarboxylase (CSD) is the rate-limiting enzyme for biosynthesis of taurine. However, the enzyme activities of taurine biosynthesis are limited in early stage of embryonic development. To analyze the expression period of TauT and CSD during embryonic development, we have investigated the gene expression of TauT and CSD using reverse transcriptase polymerase chain reaction (RT-PCR) in mouse and chicken embryos. RT-PCR anaylsis revealed that both TauT and CSD mRNAs were already expressed at Day-4.5 in mouse embryo. In chicken whole embryo, TauT and CSD mRNAs began to appear on developing times of 48 hrs and 12 hrs, respectively. TauT mRNA was detected in the organs of heart, brain and eye of the day-3 chicken embryo. Our data show that TauT and CSD mRNAs were expressed in early stage of embryonic development.

• 한국독성학회:학술대회논문집
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• 한국독성학회 2002년도 Molecular and Cellular Response to Toxic Substances
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• pp.204-204
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• 2002

Alcohol drinking during pregnancy can result in abnormal fetal development including fetal alcohol syndrome (FAS). The molecular mechanisms of FAS, however, is not completely elucidated. In the present study, we evaluated the developmental toxicity of ethanol and its metabolite, acetaldehyde using post implantation whole embryo culture and determined changes of gene expression by ethanol treatment by cDNA microarray.(omitted)

• 한국수정란이식학회지
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• 제12권2호
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• pp.181-188
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• 1997

This study was carried out to determine the effect of bovine follicular fluid(bFF), hormones, and fetal bovine serum(FBS) supplemented in the medium on the in vitro fertilization and development of bovine embryos. The ovaries were obtained from a local abattoir and placed in physiological saline kept at 30~32˚C and brought to the laboratory within 3~4 hours. The oocytes and follicular fluid were collected by aspiration from visible follicles, and the oocytes of grades I on the basis of the morphology of cumulus cells attached and the homogeneity of cytoplasmic granules were selected and used for maturation. The basal media used for oocyte maturation, fertilization and embryo development in vitro were Ham' F-10, TALP and TCM-199, respectively. The hormones supplemented in maturation medium were consisted of 35 pg /ml FSH, 10 pg /ml LH and 1 pg/mi estradiol-l7$\beta$. The bFF collected from 5~9 mm follicles was centrifuged, filtered and inactivated by heat-treatment at 56˚C for 30 min. FBS also was inactivated with the same method and kept at -20˚C until use. The embryos were co-cultured with the monolayer of bovine oviductal epithelial cells at 39˚C under 5% $CO_2$ in air for 9 days. The results obtained were summarized as follows: The fertilization rate of oocytes was found 87.4% from 10% FBS and hormones treatment for IVM, and 37.1% of these TVF embryos were developed to blastocyst stage in 10% FBS groups. Compared with this control system, the fertilization rate was decreased significantly(P<0.05) in the maturation without either FBS or hormones. These IVF embryos were developed to morula stage at the similar rate, but to blastocyst at significantly(P<0.05) lower rate in the embryo culture with or without FBS supplementation. The fertilization rate(82.9%) in hormones and 10% inactivated bFF was similar with 10% FBS and hormone groups(87.4%), but decreased significantly(P<0.05) in 20 or 30% bFF (61.0 or 66.0%), respectively. In vitro developmental competence to blastocyst stage in 10% FBS and 20% inactivated bFF(37.1% and 31.4%) was higher than in 10 or 30% inactivated bFF(20.0 or 19.2%) or 10, 20 and 30% fresh bFF(19.1, 21.0 and 17.5%) The results indicated that the in vitro fertillzation and development rate of the embryos should be improved in 10% FBS or 20% inactivated culture system and 20% inactivated bFF might be available economically for bovine oocyte maturation and embryo culture instead of fetal bovine serum.

• 한국발생생물학회지:발생과생식
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• 제18권3호
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• pp.139-143
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• 2014

Different with other fishes, the guppies (Poecilia reticulata) is ovoviviparity, which retain their fertilized eggs within the follicle throughout gestation. The synchronously growing diplotene oocytes store nutrients in droplets and yolk, before their maturation and fertilization. The lecithotrophic strategy of development entails the provisioning of embryos with resources from the maternal yolk deposit rather than from a placenta, it allows the extracorporeal culture of guppy embryo. Studies on their early development of live bearers like the guppy including lineage tracing and genetic manipulations, have been limited. Therefore, to optimize conditions of embryo in vitro culture, explanted embryos from pregnant females were incubated in embryo medium (L-15 medium, supplemented with 5, 10, 15, 20% fetal bovine serum, respectively). We investigated whether the contents of FBS in vitro culture medium impact the development of embryos, and whether they would hatch in vitro. Our study found that in 5% of FBS of the medium, although embryos developed significantly slower in vitro than in the ovary, it was impossible to exactly quantify the developmental delay in culture, due to the obvious spread in developmental stage within each batch of eggs, and embryos can only be maintained until the early-eyed. And although in culture with 20% FBS the embryos can sustain rapid development of early stage, but cannot be cultured for the entire period of their embryonic development and ultimately died. In the medium with 10% and 15% FBS, the embryos seems well developed, even some can continue to grow after follicle ruptures until it can be fed. We also observed that embryonic in these two culture conditions were significantly different in development speed, in 15% it is faster than 10%. But 10% FBS appears to be more optimizing condition than 15% one on development process of embryos and survival rate to larvae stage.

• Clinical and Experimental Reproductive Medicine
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• 제19권2호
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• pp.125-131
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• 1992

The possible effect of human follicular fluid(hFF) on the growth and development of fertilized oocytes and embryos is important because the fallopian tubes are exposed to FF after follicular rupture and the processes of fertilization and embryo cleavage occur inside the fallopian tubes. Previously, it was suggested that human FF might adversely affect on the development of early mouse embryos. In order to investigate the effect of hFF on the development of embryos, early mouse embryos were cultured in media containing various protein sources as bovine serum albumin(BSA), fetal cord serum(FCS) and FF. And we evaluated the development of early mouse embryos in terms of the morphology, cleavage rate, and cell count of blastcysts. There were no significant differences in the morula and blstocyst formation rates of 2-cell mouse embryos cultured in the media containg three different protein sources and three different concentrations of FF. The blastocyst formation rate of 1-cell mouse embryo cultured in FF group was significantly higher than that cultured in BSA group(P<0.05). The morula and blastocyst formation rates of 2-cell mouse embryos of the group cultured in the media containing FF were comparable with those of other two groups, in addition, the cell count of blastocysts of FF group in the 2-cell embryo culture was higher than those of BSA group and HCS group(P<0.01), and this finding was also noted in 1-cell embryo culture. There was no difference in the morula and blastocyst formation rates of the 2-cell mouse embryos cultured in the media containing different concentrations of FF. These results suggest that mature human follicular fluid has no inhibitory activity on the development of early mouse embryos even in high concentration and may be a good protein source which is positively associated with the development of mouse embryos in vitro especially in 1 cell embryo culture.

• 대한화장품학회지
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• 제37권3호
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• pp.247-256
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• 2011

기존의 유기계 자외선차단제는 피부투과 및 자극으로 인한 안전성의 문제가 제기되었으며, 무기계 자외선차단제는 나노화에 따른 안전성 문제가 제기되고 있다. 이로 인하여, 최근의 자외선차단제 연구는 유효성뿐만 아니라 안전성이 우수한 다양한 형태의 자외선차단제가 연구되고 있으며, 그 중의 하나가 유기-무기 결합구조의 자외선차단제에 관한 연구이다. 본 연구진은 가교된 고분자 입자 타입의 신규 자외선차단제로서 메톡시신나미도프로필실세스퀴옥산의 제조, 물성 및 유효성 평가에 대하여 보고한 바가 있다. 본 연구는 신규 자외선차단제인 메톡시신나미도프로필실세스퀴옥산의 랫드에 대한 배 태자 발생독성 연구에 관한 것으로서, 이러한 평가는 본 시험물질이 임상에서 임신 전 후에 노출 되었을 경우 불임 및 배 태아의 이상에 대한 구체적인 정보를 제공해줄 것으로 기대된다.