• Clinical and Experimental Reproductive Medicine
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• v.24 no.3
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• pp.347-353
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• 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.

• The Journal of Pediatrics of Korean Medicine
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• v.29 no.4
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• pp.97-107
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• 2015

Objectives Hataedock is the treatment that dispels toxic heat and meconium gathered at the fetus for the new born baby by orally administered herbal extracts. The purpose of this study was to evaluate whether Hataedock alleviate inflammatory skin damages in AD-induced NC/Nga mice through regulating of skin barrier maintain and Th2 differentiation. Methods We established an AD model in the 3-week-old NC/Nga mice through the repeated application of DNFB (dinitrochlorobenzene) on days 28, 35, 42 after Hataedock treatment which was orally administered. We identified the changes of skin barrier and Th2 differentiation through the histological and immunohistochemical changes of protein kinase C (PKC), interleukin (IL)-4, degranulated mast cell, Substance P and MMP-9. Results Our results suggested that Hataedock treatment significantly down-regulated levels of PKC by 82% (p < 0.001), as well as IL-4 by 56% (p < 0.001). Hataedock also suppressed mast cell infiltration, ear edema formation. and Substance P in the tissue of NC/Nga mice were decreased by 57% (p < 0.001), and MMP-9 by 55% (p < 0.001). Conclusions These results suggest that Hataedock alleviates AD through the down-regulation of PKC and Th2 cytokines, which are involved in the initial steps of AD development. Hataedock have potential application for the treatment of AD.

• The Journal of Pediatrics of Korean Medicine
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• v.31 no.3
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• pp.37-45
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• 2017

Objectives Hataedock is an orally-administered herbal extract that used to treat toxic heat and help excrete meconium accumulates in the fetus from pregnancy. This study compared the effects of Hataedock using the extract of Douchi and omega-3 to control inflammation in the colonic mucosa. Methods NC/Nga mice were divided into three groups; Control group (no treatment), FGT group (given the extract of Douchi), and O3T group (given omega-3). The 3-week-old mouse from each group were given the extract of Douchi and omega-3. After 2 weeks, the colonic mucosa tissues of the mice were observed via Immunohistochemical staining for IL-4, IL-13, CD40, $Fc{\varepsilon}RI$, $p-I{\kappa}B$, EGF in the intestinal mucosa. Results In both FGT and O3T group, IL-4, IL-13, STAT-6 levels and CD40-positive reactions were decreased. However, EGF-positive responses were increased. FGT group showed 35% decrease in IL-4, 45% decrease in IL-13, 48% decrease in CD40, 50% decrease in $Fc{\varepsilon}RI$, 54% decrease in $p-I{\kappa}B$, 52% increase in EGF compared to the control group. O3T group also showed 14% decrease in IL-4, 23% decrease in IL-13, 22% decrease in CD40, 29% decrease in $Fc{\varepsilon}RI$, 41% decrease in $p-I{\kappa}B$, 39% increase in EGF compared to the control group. Conclusions Both early administration of Douchi extract and omega-3 were effective in regulating Th2, relieving inflammation and developing colonic mucosal tissues. Hataedock with Douchi extract may be more effective in modulating immune system in colonic mucosa than omega-3.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.23 no.3
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• pp.205-211
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• 2001

Cleft palate is one of the most serious congenital anomalies in human that causes a sucking problem in newborn babies and morphologic deformity that usually leads to death in newborn mouse offspring due to an insufficient ability to suck milk. Therefore cleft palate had been researched with epidemiologic and molecular methods, and many etiologic factors were examined closely. Among of the research methods, biologic molecule researches have been more important method for cleft palate formation study. The $TGF-{\beta}$ had an important role in the cell migration, epithelial-mesenchymal transdifferentiation, extracellular matrix synthesis and deposition. But there was a little research which was study about correlation cleft palate induced by beta-aminonitroproprionitrile(BAPN) with $TGF-{\beta}$ expression. A purpose of this presented study was examed how $TGF-{\beta}$ expression in cleft palate mice. At gestation days 13, BAPN-monofumarate salts($(C_3H_6N_2)_2$ ${\cdot}$ $C_4H_4O_4$, Sigma Co.) was single oral administered to 4 pregnant rats according to 1g/kg body weight. And pregnant rats were sacrificed on day 20 post coitus(p.c.), The $TGF-{\beta}$ expression patterns of cleft formed fetus mice was followed that; 1.Osteoblast, mesenchymal cell and epithelial cell of cleft mice were low expression compare to control mice. 2.There was no $TGF-{\beta}$ difference expression pattern of osteocyte of cleft mice compare to control mice. 3. In western blot analysis, thickness of band of $TGF-{\beta}$ in cleft mice was thin and dilute compare to control mice.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.34-36
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• 2002

Epidermal growth factor (EGF) and insulin-like growth factor-I (IGF-I) have been shown to stimulate proliferation and differentiation of various somatic cells, including placental trophoblasts and also to enhance fetal growth and development when maternally administered. Since an increase of the expression of placental EGF and IGF-I receptors in rat, mouse, and human with the gestation advanced, both EGF and IGF-I were considered to play pivotal roles on fetal growth by regulating some function of placental cells. Amino acids are crucial importance for both maternal and fetal requirements of energy source and essential constituent of fetal mass during pregnancy. Impaired fetal and placental uptake of amino acids has been observed in several models of growth retardation in the rat. Amino acid is concentrated in the fetal side through active transport by amino acid transporters and is one of the important metabolic fuels for the fatal growth. Therefore, at first plasma amino acid concentrations in mothers and fetuses were measured as an index of uphill transport across the placenta associated with EGF and IGF-1. The EGF administration at the concentration of 0, 0.1, or 0.2 $\mu\textrm{g}$/g to pregnant rats from day 18 to 21 of gestation apparently increased fetal/maternal ratio of serum proline concentration and also fatal growth in EGF dose-dependent manner. When IGF-I in doses of 0, 1, 2, and 4 $\mu\textrm{g}$/g were administrated, the ratio of leucine, isoleucine, tryptophan, phenylalanine, tyrosine and also fetal growth significantly increased with a dose-dependent manner. These results suggested that EGF and IGF-I enhanced fatal growth by, as one of its possible mechanisms, promoting placental activity to transfer some amino acid supplies from the mother to the fetus in late pregnancy.

• Korean Journal of Veterinary Research
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• v.40 no.1
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• pp.42-48
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• 2000

Akabane disease is transmitted through mosquitoes in cattle, sheep and goats. It shows congenital abnormalities including encephalomyetitis, hydranencephaly, neurogenic arthrogryposis, and deformed neonatal calves. Akabane viruses, 93FMX and K-9 strain, were isolated from fetal matrix of aborted cow and blood of healthy cow, respectively. S gene sequences of 93FMX and K-9 showed 100% homology with that of OBE-1 strain isolated in Japan. Based upon our sequencing data, we synthesized specific primers for PCR diagnosis. Using these primers, we were able to amplify the S gene of Akabane virus not only from the culture fluid of Vero cells but also from the brain tissue of suckling mouse inoculated with, Akabane virus. These PCR products were confirmed by Southern blot hybridization. Not only the sensitivity of PCR test was high enough to detect the viruses of $10^{1.0}TCID_{50}/ml$, but also the time for diagnosis was significantly shorter than that of the virus isolation by tissue culture method. This method was also effective for the detection of Akabane virus in the cerebrum of fetus. RT-PCR method may be used for a useful diagnostic test of the clinical cases of Akabane disease.

• Korean Journal of Agricultural Science
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• v.49 no.1
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• pp.11-18
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• 2022

A disrupted immune system during pregnancy is involved in pregnancy complications, such as spontaneous abortion, preeclampsia, and recurrent pregnancy loss. This study examined the role of toll-like receptor (TLR) 4 and ASC (apoptosis-associated speck-like protein containing a CARD [c-terminal caspase recruitment domain]) in pregnancy complications using a lipopolysaccharide (LPS)-induced miscarriage mice model. Incidences of miscarriage and embryonic resorption were examined at 9.5 days of pregnancy in wild-type (WT), ASC knockout (KO), and TLR4 KO mice after injecting them with LPS. The fetuses and placenta were obtained after sacrifice at 15.5 days of pregnancy. A significantly lower frequency of fetus absorption was found in TLR4 KO mice, whereas corresponding absorption outcomes were strongly induced in the WT and ASC KO mice upon an LPS injection. As expected, TLR4 KO mice were resistant to LPS-induced abortion. A histological analysis of the miscarried placenta showed increasing levels of the eosin staining of spongiotrophoblast cells without any obvious difference between WT and ASC KO mice. These results suggest that TLR4 KO mice are resistant to LPS, which affects pregnancy persistence, whereas WT and ASC KO mice show high miscarriage rates due to LPS. Moreover, the ASC adaptor is not directly involved in LPS-induced miscarriages, and the NLRP3 inflammasome can be activated by other proteins in the absence of ASC.

• Clinical and Experimental Reproductive Medicine
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• v.24 no.2
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• pp.261-265
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• 1997

The objective of this study was to examine the effect of EGF to the in vivo development of mouse IVF/IVC embryo. The 2-cell embryos were cultured in medium (5-6 embryos/25 ${\mu}l$/drop) w/wo EGF (10 ng/ml) and day 4 blastocysts recovered from each treatment group were transferred into the uteri of recipients of pseudopregnant day 3. The results obtained in this experiment were summarized as follows: 1. When the effect of EGF to the in vitro development and cell number of blastocysts produced from the culture of 2-cell embryos in w/wo EGF was determined, those results of EGF treatment group showed not significant difference compared with control. 2. However, when the effect of EGF to the in vivo development of blastocysts recovered from each treatment group was examined, production of the normal fetus against transferred embryos in EGF treatment group (51.2%) was very higher than that in control group (31.1%), although total implantation was not significantly different between treatment group (control: 64.4%, EGF: 69.8%). Therefore, this result suggested that EGF can affect to the in vivo development of IVF/IVC embryos through the improvement of embryo quality, although EGF treated embryos showed not significant development rate compared with control.

• Animal Bioscience
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• v.36 no.1
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• pp.43-52
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• 2023

Objective: This study aimed to examine the influence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection on pregnancy in cytokeratin-18 (K18)-hACE2 transgenic mice. Methods: To determine the expression of hACE2 mRNA in the female reproductive tract of K18-hACE2 mice, real-time polymerase chain reaction (RT-PCR) was performed using the ovary, oviduct, uterus, umbilical cord, and placenta. SARS-CoV-2 was inoculated intranasally (30 μL/mouse, 1×10⁴ TCID₅₀/mL) to plug-checked K18-hACE2 homozygous female mice at the pre-and post-implantation stages at 2.5 days post-coitum (dpc) and 15.5 dpc, respectively. The number of implantation sites was checked at 7.5 dpc, and the number of normally born pups was investigated at 20.5 dpc. Pregnancy outcomes, including implantation and childbirth, were confirmed by comparison with the non-infected group. Tissues of infected mice were collected at 7.5 dpc and 19.5 dpc to confirm the SARS-CoV-2 infection. The infection was identified by performing RT-PCR on the infected tissues and comparing them to the non-infected tissues. Results: hACE2 mRNA expression was confirmed in the female reproductive tract of the K18-hACE2 mice. Compared to the non-infected group, no significant difference in the number of implantation sites or normally born pups was found in the infected group. SARS-CoV-2 infection was detected in the lungs but not in the female reproductive system of infected K18-hACE2 mice. Conclusion: In K18-hACE2 mice, intranasal infection with SARS-CoV-2 did not induce implantation failure, preterm labor, or miscarriage. Although the viral infection was not detected in the uterus, placenta, or fetus, the infection of the lungs could induce problems in the reproductive system. However, lung infections were not related to pregnancy outcomes.

• Korean Journal of Animal Reproduction
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• v.18 no.4
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• pp.299-307
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• 1995

The goal of cell stem cell technology is to produce a viable and genetically normal animal. To achieve this goal various laboratories have followed 2 different pathways beginning with either the culture of 1) single or pooled ICMs grown with or without a feeder layer or 2) single or pooled 16-20 cell stage embryos grown with a feeder layer. Also, thus far embryonic cell cultures or lines have been established by several methods including loose suspension culture for short-term cultures and more commonly murine or bovine fibroblast feeder layers for long-term culture. Pluripotent lines have been derived from 16-cell through blastocyst inner cell mass stages. The efficiency of establishing cell lines and cell proliferation apper to be affected by the number of cells or embryos starting the line. Most attempts to produce offspring from long term STO cell feeder layer cultured ICM or morulae derived ES cells have resulted in pregnancy failure in the first trimester when ES cells were used in cuclear transfer or have failed to retain ES cells in the progeny produced by chimerization. The exception is 1 chimeric fetus from use of morula ES cells in the chimerization with early embryonic cells. There is much to be learned yet about ES cell culture requirements for maintenance of totipotency. If bovine ES cell lines loose imprinting pattern and totipotency with long-term culture and passage as suggested for mouse ES cells, we may be limited to the use of short-term cultures for multiplication of embryos and efficient production of transgenic animals. No bovine ES cell system has yet met all of the criteria indicated for a totipotent ES cell line.