Chung, H. M.;Lee, K. S.;Kim, J. U.;Lim, J. M.;Song, B. K.;Chang, J. B.;Ko, J. J.;Yoon, T. K.;Park, Chan;Cha, K. Y.
Journal of Embryo Transfer
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v.15
no.1
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pp.9-14
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2000
This experiment was undertaken to investigate the effect of administration of Agyoju or Mokhyang on the maintenance of pregnancy, delivery and sex ratio in the mice in different gestation periods. Adult female mice were administered orally at three different periods, from ovulation to implantation (Exp.1), from post-implantation to organogenesis (Exp.2), and from fetal growth to parturition (Exp.3). In Experiment 1, number of fetus implanted and mean body weight were not significantly different. However, the delivery of male offspring was significantly increased in Agyoju and Mokhyang administrated groups than control group. In Experiment 2, the number of fetuses implanted, live offsprins and their body weight at delivery were significantly increased in the Agyoju administered group than Mokhyang and control groups. In Experiment 3, the number of live offspring and sex ratio were not different in both treatments and control group. However, mean body weight at delivery was significantly increased in both treatment groups than that of control group. These results suggest that 1) Agyoju and Mokhyang have beneficial effects in maintenance of pregnacy, and that 2) The action of unknown component(s) in Agyoju may be related to selection of male spermatozoa for fertilization in vivo, and that 3) the administration of Agyju of Mojhyang during mid-and late-pregnance periods were shown the increment of body weight of live offspring without decrease of litter size.
The objective of this study was to investigate the in vitro / in vivo embryonic development after vitrification of mouse zygotes and the chrom osomal normality of delivered live young after embryo transfer. Mouse IVF zygotes were cryopreserved by vitrification using vitrification solution, EFS40 (40% ethylene glyc이, 30% Ficoll a and 0.3 M sucrose in phosphate buffer saline c containing 10% FBS ) . After mouse zygotes were exposed to EFS40 at 25"C for 30 sec., they were immediately plunged into LN$_2$. Vitrified thawed mouse zygotes were cultured upto bIastocysts in M16 for 4 days. The rates of in vitro development were 71.5% under this condition. Cultured blastocysts were transferred to recipients (3 day of pseudopregnant) on one or both uterus horns (6-8 embryos per a uterus horn). And all recipients were allowed to produce litters. The results obtained in these experiments were summarized as follows: The pregnancy rates and in vivo survival rates, live fetus rates, for vitrified zygotes (80.0, 39.6%) were not significantly difference in those of control zygotes (77.8%, 50.0%). Also, all of live-born young mice were chromosomally normal (n=40). This results suggested that proposed rapid vitrification procedures can be effectively use in 1-cell mouse zygotes cryopreservation.
Kim, M.K.;Kim, E.Y.;Yi, B.K.;Yoon, S.H.;Park, S.P.;Chung, K.S.;Lim, J.H.
Clinical and Experimental Reproductive Medicine
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v.24
no.3
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pp.347-353
/
1997
This study was to test whether in vitro/in vivo survival of vitrified mouse blastocysts was influenced by culture conditions and ET method. Mouse blastocysts were obtained from in vitro fertilization and cultured for 4 days in M16 medium, and they were vitrified in EFS40 which contained 40% ethlyene glycol, 18% Ficoll and 0.5 mol sucrose in PBS. In experiment I, in vitro and in vivo survival rate of these embryos were evaluated in different culture condition after thawing. When thawed embryos were cultured in M16 medium as a control, m-CR1 medium contained 20 amino acids (2% BME amino acis and 1% MEM non-essential amino acids solution) and 4 mg/ml BSA and cumulus monolayer cell co-cultured condition in mCR1 medium (10% FBS), their in vitro survival at 24 hr after thawing was not affected by culture condition (75.6, 83.1, 82.4%). However, in vivo survival rates of implantation in m-CR1 medium (80.4%) were significantly higher than those of M16 medium (51.2%), co-culture (57.1%) condition, although there was no difference in live fetuses rates on day 15 gestation (39.0, 49.0, 38.1%). In experiment II, the in vivo development potential of embryos by ET methods was examined. When blastocysts were transferred to the day 2, 3 pseudopregnant recipient without culture soon after thawing, no pregnant recipient was obtained on the day 2 pseudopregnancy, and 50% of pregnancy rates and 15.4% of live fetus rates were obtained on the day 3 pseudopregnant recipients. These results were significantly lower than those of transferred group (day 3 pseudopregnant recipients) after culture for 16 hr post thawing (73.5, 57.1%) (p<0.05). In experiment III, to elevate usability of delayed embryos in vitro/in vivo survival of vitrified embryos (day 4 early, day 5 early and expanding blastocyst) were examined. in vivo survival rates (live fetus, total implantation) were higher in day 4 early blastocysts (33.3, 66.7%) than in day 5 expanding blastocysts (29.0, 38.7%), although the highest in vitro survival rates were obtained in the day 5 expanding brastocysts (78.3%). Therefore, these results suggest that the in vitro/in vivo survival rates of vitrified embryos could be improve by the culture condition and ET method and that the in vivo development rates of delayed embryos were decreased with longer culture duration in vitro. It means that more effective cryopreservation was obtained in day 4 early blastocysts than in day 5 expanding blastocysts.
Reproductive traits of sows are determined chiefly by the genetic background of the dam herself. Whether the breed of the serviced boars also contributes is not yet clear. A total of 8,946 litters of Landrace (L) sows from 68 farms were examined. Of these, 876, 1,260, and 6,810 litters were produced from natural mating with L, Yorkshire (Y), and Duroc (D) breeds, respectively. The birth litter size (BLS), weaning litter size (WLS), live born piglets (LBP), and survival rate % (SR), of every litter were recorded. A general linear-model procedure was followed to estimate the effects of boars' breed (B), parity (P) and B${\times}$P interaction on the reproductive traits of the sows. Results show that BLS, WLS, and LBP were all significantly (p<0.001) affected by B and P. SR% was significantly influenced by B but not by P. L sows crossbred with Y or purebred with L produced litters with higher BLS, WLS, and LBP values than those bred with D. Pure L breeding yielded litters with lower SR% than did crossbreeding with D, while the difference between SR% due to LY and that due to LD crossbreeding was not significant. The interaction of B with P was significant with respect to BLS, WLS, and LBP (p<0.001), but not SR%. No significant B effect on reproductive traits was measured in sows at their first parity; but at latter parities, LL or LY produced litters with similarly high BLS, WLS and LBP, which values were all significantly greater than those of LD litters. The breed of boar evidently affected the subsequent reproductive performance of L sows and this effect may be further manipulated by the parity effect. Breed differences in semen quality and the success of fetus development with different interactions of the genetic background with the uterus function of the sow that may contribute to these effects are discussed.
Proceedings of the Korean Society of Developmental Biology Conference
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2003.10a
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pp.7-8
/
2003
Since it was first reported in 1997, somatic cell cloning has been demonstrated in several other mammalian species. On the mouse, it can be cloned from embryonic stem (ES) cells, fetus-derived cells, and adult-derived cells, both male and female. While cloning efficiencies range from 0 to 20%, rates of just 1-2% are typical (i.e. one or two live offspring per one hundred initial embryos). Recently, abnormalities in mice cloned from somatic cells have been reported, such as abnormal gene expression in embryo (Boiani et al., 2001, Bortvin et al., 2003), abnormal placenta (Wakayama and Yanagimachi 1999), obesity (Tamashiro et ai, 2000, 2002) or early death (Ogonuki et al., 2002). Such abnormalities notwithstanding, success in generating cloned offspring has opened new avenues of investigation and provides a valuable tool that basic research scientists have employed to study complex processes such as genomic reprogramming, imprinting and embryonic development. On the other hand, mouse ES cell lines can also be generated from adult somatic cells via nuclear transfer. These 'ntES cells' are capable of differentiation into an extensive variety of cell types in vitro, as well assperm and oocytes in vivo. Interestingly, the establish rate of ntES cell line from cloned blastocyst is much higher than the success rate of cloned mouse. It is also possible to make cloned mice from ntES cell nuclei as donor, but this serial nuclear transfer method could not improved the cloning efficiency. Might be ntES cell has both character between ES cell and somatic cell. A number of potential agricultural and clinical applications are also are being explored, including the reproductive cloning of farm animals and therapeutic cloning for human cell, tissue, and organ replacement. This talk seeks to describe both the relationship between nucleus donor cell type and cloning success rate, and methods for establishing ntES cell lines. (중략)
In differentiating human embryonic stem (d-hES) cells there are a number of types of cells which may secrete various nutrients and helpful materials for pre-implantation embryonic development. This study examined whether the d-hES could function as a feeder cell in vitro to support mouse embryonic development. By RT-PCR analysis, the d-hES cells revealed high expression of three germ-layered differentiation markers while having markedly reduced expression of stem cell markers. Also, in d-hES cells, LIF expression in embryo implantation-related material was confirmed at a similar level to undifferentiated ES cells. When mouse 2PN embryos were cultured in control M16 medium, co-culture control CR1aa medium or co-cultured with d-hES cells, their blastocyst development rate at embryonic day 4 (83.9%) were significantly better in the d-hES cell group than in the CR1aa group (66.0%), while not better than in the M16 group (90.7%)(p<0.05). However, at embryonic days 5 and 6, embryo hatching and hatched-out rates of the dhES cell group (53.6 and 48.2%, respectively) were superior to those of the M16 group (40.7 and 40.7%, respectively). At embryonic day 4, blastocysts of the d-hES cell group were transferred into pseudo-pregnant recipients, and pregnancy rate (75.0%) was very high compared to the other groups (M16, 57.1%; CR1aa, 37.5%). In addition, embryo implantation (55.9%) and live fetus rate (38.2%) of the d-hES cell group were also better than those of the other groups (M16, 36.7 and 18.3%, respectively; CR1aa, 23.2 and 8.7%, respectively). These results demonstrated that d-hES cells can be used as a feeder cell for enhancing in vitro and in vivo developmental potential of mouse pre-implantation embryos.
The adverse effect of diving on the fetus may extend beyond n gestational process and outcome. Primiparous Sprague-Dawley rats were assigned to one of ten exposure schedules during gestatred $PO_2$ level, the following question about the effect of exposing a pregnant female to high partial pressure of inspired oxygen has been raised. 'What effect does an increased maternal $PIO_2$ have on fetal arterial $PO_2$ and therefore on possible fetal oxygen poisoning?' This study was carried out to observe the effects of maternal hyperoxia on gestational process and outcome. Primiparous Sprague-Dawley rats were assigned to one of ten exposure schedules during gestation. The treatment groups were subjected to either the high concentration of oxygen, or the high atmospheric pressure. On day 21 of gestation, laparotomy was performed to examine for number and distribution of implantations and live and resorbing embryos. Fetuses were weighed, and examined for gross malformations. Subsequently, they were fixed, measured in physical parameters, and examined for visceral anomalies. Minor visceral anomalies and anatomical variation was not found. Similarily, there were no significant differences when number of resorptions, mean fetal weights, pregnancy interruption rate were compared by analysis of variance. These results indicate that exposing rats to oxygen at increased atmospheric pressure doese not affect fetal health or survival.
The purpose of this study was to find out the factors affecting birth sex ratio which had revealed alarmingly higher in recent years. This study was conducted to get hold of the basic data such as the sex ratio of live births which had been delivered at 5 general hospitals in Taegu from 1982 to 1986. And author chose Dongsan Medical Center from those 5 hospitals for further detailed study, and reviewed 1,286 medical records of mothers who had birth in each February and March from 1984 to 1987. Of these 1,286 deliveries, 30 cases were twin deliveries, which added the total children numbers to 1,316. The findings of this study were as follows: The average of birth sex ratio( 114.5) for 5 years at five general hospitals in Taegu was slightly higher than the traditional birth sex ratio( 102-107) and the highest was 123.4 in 1985. There was no significant difference in birth sex ratio by mother's educational level, her religion nor father's occupation. The birth sex ratio of the cases whose mother was above 35 years old was significantly high(400.0), that of the cases who were born to the mothers who had more than 3rd parity was significantly high(309.5), and that of the cases whose mother had more than two daughters and no son was also significantly high (330.7). The birth sex ratio of the cases who did not have ultrasonogram on their prenatal visits was 87.8. In contrast to this, that of the cases who had ultrasonogram was significantly higher( 135.5). Of the mothers who have more than two daughters and no son' 80.4% (45 cases) had ultrasonogram and their newborn's birth sex ratio was significantly high(542.9). Of the cases who were born to more than 3rd parity mothers, 75.6%(65 cases) of their mothers had ultrasonogram and their sex ratio was significantly high(441.6). It was revealed by this study that the birth sex ratio is changed to significantly high due to sex confirmation of the fetus by ultrasonogram. Thus it is recommended that the value related to having equality of sexes must be ingrained by changing the various social systems and value systems which is now aggrevating the son preference and should provide controlling system which can prohibit the ultrasonogram for sex confirmation only.
Kim, Chang-Seok;Lee, Sun-Dong;Kim, Pan-Gyi;Lee, Jang-Woo;Park, Hae-Mo
Journal of Society of Preventive Korean Medicine
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v.9
no.2
/
pp.59-75
/
2005
The experiments were undertaken to evaluate the effects of Bosaengtang in pregnant rats and fetuses. Female Sprague-Dawley rats were orally administered with Bosaengtang at the dose of 5mg/kg/day for 20 days. Pregnant rats were sacrificed at the 20th day of gestation, and observed internal and reproductive organs. Fetuses were randomly selected and fixed in 95% ethanol. Fetuses were stained with alcian blue and alizarin red S, and observed skeletal malformations. The results obtained were as follows : Bosaengtang administered group showed higher maternal body weight than the control group, but both groups showed increase in weight. Bosaengtang administered group showed lower than the control group, and higher liver and kidney weight than the control group, but the differences were minimal. There were no significant changes between the control and treated group in blood chemistry values and hematological values but all the groups were within in normal ranges. There were no significant changes in the number of corpus luteum, implantation, live fetus and implantation rate, delivery rate, late resorption rate, sex ratio, but Bosaengtang administered group showed higher early resorption rate than control group. comparing the control and Bosaengtang group, neonatal body weight and the number of fetuses were increased in Bosaengtang group. The fetuses of dams treated with Oriental medicine didn't showed external malformation. Vertebral and sternal variations were observed in Bosaengtang group, but the differences were not apparent compared to the control group. The number of ribs, cervical, thoracic and lumbar vertebrae were normal. The number of sacral was similar and the number of caudal was increased. Fetuses showed significant difference in the number of caudal vertebrae. (P<0.01) From these results, we can carefully conclude that Bosaengtang showed beneficial effects on maternal body weight, early resorption rate, number of live fetus. There were no significant changes in organ weight, hematoscopy, reproduction organs. External malformation wasn't visible. Skeletal variations were showed in vertebrae and sternum but compared to the control group, these variations weren't much different.
Rubella is a viral disease with mild constitutional symptoms and generalized rashes. In childhood, it is an inconsequential illness, but when it occurs during early pregnant period, there are significant risks of heart defects, cataract, mental retardation to the fetus. The series of congenital defects induced by rubella is called 'congenital rubella syndrome'. Many research have been performed to find out more effective prevention program on rubella. The objectives of this study are, first, to calculate the incidence rate of acute rubella infection and congenital rubella syndrome in Korea, second, to evaluate economic efficiency of several rubella vaccination policies and to offer data for the most reasonable decision on vaccination policy. Study populations are 663,312 children of one year-old in 1992. The author has performed cost-benefit analyses according to the three vaccination policies-U.S.A.'s. U.K.'s and Sweden's. In this Study, the author got the incidence rate of acute rubella infection using the catalytic model. In the meantime, the author used 50 per 100,000 live births as the incidence rate of congenital rubella syndrome. The discount rate used in this study was 5 percent per annum. The sensitivity analyses were done with different discount rates (4%, 7%) and different incidence rate of congenital rubella syndrome (10,100 per 100,000 live births) : The study results are as follows: 1. Without vaccination, lifetime expenditures per patient for acute rubella infeciton amount to 14,822 won and the total expenditures to about 3.1 billion won. Meanwhile, lifetime expenditures per patient for congenital rubella syndrome amount to about 91 million won and the total expenditures to about 16.3 billion won without vaccination. 2. The cost of vaccination for a child of one year old was 2,322 won and the total cost for the one year old children was about 1.5 billion won (American style). The cost for vaccination of female children at fifteen was about 339 million won (Birtish style). And the cost of vaccination at one for both sex and female children at fifteen was about 1.9 billion won (Swedish style). 3. The benefit to cost ratios of vaccination of female children at fifteen that is the british mode of rubella vaccination, was 60.0 at the level of 80% population coverage and 48.6 at 100% coverage. It shows much higher benefit to cost ratio than those of the other two vaccination policies. 4. Both net benefits of vaccination at one (American style) and that of vaccinations at one and fifteen (Swedish style) range from about 17.0 billion to 17.8 billion won, those were larger than that of vaccinations of female children at fifteen (Birtish style, about 16.0 billion). 5. In marginal cost-benefit analysis of only additional program of revaccination, the benefit to cost ratios were 3.6 (80% coverage rate) or 0.6 (100% coverage rate). It implies that additional program was less efficient or inefficient. 6. In sensitivity analysis with different discount rates(4% or 7%) and different incidence rates of congenital rubella syndrome (10 or 100 per 100,000 live births), the benefit to cost ratios has fluctuated in wide range. However, all the ratios of vaccination of female children at fifteen were higher than those of the others. Even under the most conservative assumption, the benefit to cost ratios of all the rubella vaccination policies were higher than 3.3. In conclusion, all the rubella vaccination policies found to be cost-effective and particularly the vaccination of female children at fifteen was strongly recommended.
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