• Title/Summary/Keyword: in vitro study

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Mouse Embryo Culture used in Quality Control of Water for Human in Vitro Fertilization : The One-cell Stage Versus the Two-cell Stage Model (수질에 대한 1-세포기 및 2-세포기 생쥐배아를 이용한 생물학적 정도관리에 관한 연구)

  • Lee, Ye-Kyung;Chung, Hye-Won;Kim, Hyung-Mee;Oh, Seung-Eun;Son, Young-Soo;Yu, Han-Ki;Woo, Bock-Hee
    • Clinical and Experimental Reproductive Medicine
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    • v.20 no.1
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    • pp.9-17
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    • 1993
  • This study was carried out investigate the effect of water quality and the kind of media on the in vitro development of 1-cell and stage mouse embryos. $F_1$ hybrid mice were superovulated and timely mated. 1-cell stage and 2-cell stage mouse embryos were recruited and taken into Ham's F-10 or m-KRB media which was made of two of two kinds of water having different quality, highly purified water and tap water. 2-cell stage embryos grew up well in vitro to blastocyst or hatching blastocyst regardless of the composition of culture media, but 1-cell stage mouse embryo didn't develop well to blastocyst or hatching blastocyst in simple media like m-KRB. These results meant in vitro devleopment of 1-cell stage mouse embryo neded complex media like Ham's F-10 which contained abundant protein components. In case of quality control for water, in vitro fertilization program. observation of in vitro development of 2-cell mouse embryos up to blastocyst or hatching blastocyst media such as m-KRB would be efficatious in detecting the difference of water quality.

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Study on the In-vitro Culture Method for Normal Embryonic Cell Development of Porcine Parthenogenetic Embryos

  • Jung, Na-Hyeon;Kim, Sang-Hwan;Kim, Dae-Seung;Yoon, Jong-Taek
    • Journal of Animal Reproduction and Biotechnology
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    • v.35 no.1
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    • pp.94-101
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    • 2020
  • In the early development of parthenogenetic embryo, cytoplasm and nucleic acid fragmentation may be a cause of lower embryo development. The purpose of this study was to evaluate whether embryonic development and apoptosis factors can be reduced by controlling the in-vitro culture environment by the addition of hormones, pregnancy serum and uterine milk. Our study showed that the activity of Casp-3 increased within the cytoplasm when artificially used hormones to induce the incubation environment, and PCNA's manifestation was low. However, the addition of pregnant serum appeared to lower the Casp-3 activity compared to the other groups. In addition, MMP-9 activity was increased and early embryo development and cytoplasmic fidelity were also increased. Therefore, the results of the present study showed that the use of gestational serum in the development of parthenogenetic embryo inhibit apoptosis and increases cytoplasmic reorganization by natural environmental control in in vitro culture.

Changes of the Microbial Population and Determination of Pepsin and In vitro Digestibilities of Pasteurized and Cured Food Wastes (살균${\cdot}$숙성된 남은 음식물의 공정별 미생물 분포 및 Pepsin과 In vitro 소화율 평가)

  • Baik, Y. H.;Ji, K. S.;Kwak, W. S.
    • Journal of Animal Environmental Science
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    • v.11 no.2
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    • pp.125-134
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    • 2005
  • This study was conducted to evaluate changes of microbial population, pepsin digestibility of protein and in vitro digestibility of nutrients of food waste mixture pasteurized and cured using a rotary drum system. A pasteurization process (30 min at $80^{\circ}C$) tended to decrease microbial populations and eliminated (P<0.05) molds in food waste mixture. The subsequent curing process increased (P<0.05) lactic acid bacteria counts which were reduced by the heated pasteurization process. The heated pasteurization process decreased (P<0.05) pepsin digestibility of protein in food waste mixture. In vitro digestibilities of dry matter and organic matter were high in the order of bakery by-product, wheat bran, food waste (=barley bran). These results indicate that food waste mixture pasteurized and cured using a semi-dehydration rotary drum system may be an effective animal feed resource.

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Normal and Abnormal Fertilisation of Zebu Cattle Oocytes In Vitro

  • Talukder, Anup Kumar;Shamsuddin, Mohammed;Rahman, Mohammad Bozlur;Bari, Farida Yeasmin;Parish, John J
    • Journal of Embryo Transfer
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    • v.24 no.2
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    • pp.89-95
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    • 2009
  • Successful in vitro embryo production heavily relies on the normal maturation and fertilisation of oocytes. We examined the normal and abnormal fertilisation of zebu cattle oocytes matured in vitro. Immature cumulus oocyte complexes (COCs) from zebu cattle ovaries at slaughter were matured in vitro (IVM) for 24 h. The oocytes were either fixed, stained and examined for nuclear changes or fertilised in vitro (IVF) with Percoll-separated, heparintreated spermatozoa (1.0 ${\times}$ $10^6$/mL) of zebu (n = 7) and crossbred bulls (n = 7). After 18 h of sperm-COCs co-incubation at $39^{\circ}$C with 5% $CO_2$ in humidified air, the presumptive zygotes were fixed, stained and examined for pronuclei. The number of oocytes retrieved per ovary was 5.4 ${\pm}$ 0.7. The percentage of matured oocytes was 73.0. The difference in motility of spermatozoa before and after Percoll seperation was significant (p<0.001). The percentages of normal and abnormal fertilisation (polyspermia and oocytes with one pronucleus) varied significantly depending on individual bulls (p<0.05). A protocol for IVF of IVM oocytes in Bangladeshi zebu cattle is developed. A future study may elucidate the capacity of such IVM-IVF oocytes to develop to the blastocyst stage for transfer to surrogate mother.

Characterization and Preparation of Low Molecular Weight Water Soluble Chitosan Nanoparticle Modified with Cell Targeting Ligand for Efficient Gene Delivery (효과적인 유전자전달을 위한 표적성 리간드가 도입된 저분자량 수용성 키토산 나노입자의 제조 및 특성)

  • Heo, Sun-Heang;Jang, Min-Ja;Kim, Dong-Gon;Jeong, Young-Il;Jang, Mi-Kyeong;Nah, Jae-Woon
    • Polymer(Korea)
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    • v.31 no.5
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    • pp.454-459
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    • 2007
  • Gene therapy using low molecular weight water soluble chitosan (LMWSC) as polycationic polymer shows good biocompatibility, but low transfection efficiency. The mechanism of folic acid (FA) uptake in the cells to promote targeting and internalization could improve transfection rates. The objective of this study was to synthesize and characterize the WSCFA-DNA complex and evaluate their cytotoxicity, in vitro. In $^1H-NMR$ spectra, specific peaks appeared both of FA and LMWSC in $D_2O$. WSCFA nanoparticles have spherical shapes with particle size show below 110 nm. In the cell cytotoxicity test, the WSCFA-DNA complex showed high cell viability, in vitro. Gel electrophoresis showed condensed DNA within the carriers. hi vitro transfection efficiency was assayed by fluorescence spectroscopy WSCFA nanoparticles have less cytotoxicity, good DNA condensation and particle size around 110 nm, which makes them a promising candidate as a non-viral gene vector.

In vitro and in vivo antibacterial activity of Meliae fructus extract against Helicobacter pylori (Helicobacter pylori에 대한 천련자 추출물의 in vitro와 in vivo 실험에서의 항균활성)

  • Lee, Hyun-A;Kim, Okjin
    • Korean Journal of Veterinary Research
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    • v.52 no.2
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    • pp.105-113
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    • 2012
  • In this study, a medicinal herbal plant, Meliae fructus, was examined and screened for anti-Helicobacter (H.) pylori activity. Seventy percent ethanol was used for herbal extraction. For anti-H. pylori activity screening, inhibitory zone tests as an in vitro assay and in vivo study using a Mongolian gerbil (Meriones unguiculatus) model were performed. Also, the safety of herbal compounds was evaluated by animal study. As a result of inhibitory zone test, Meliae fructus extract demonstrated strong anti-H. pylori activities. Also, as results of in vivo animal studies, Meliae fructus demonstrated strong therapeutic effects against H. pylori infection according to the criteria of histological examination and rapid urease test. As results of the safety study, after 28 days treatment of the Meliae fructus extract, the animals were not detected any grossly and histological changes. These results demonstrate that it can be successfully cured against H. pylori infection and protected from H. pylori-induced pathology with Meliae fructus. It could be a promising native herbal treatment for patients with gastric complaints including gastric ulcer caused by H. pylori.

Hepatoprotective activity of terpenoids and terpenoid fractions of Scoparia dulcis L

  • Krishnamurthy, Praveen Thaggikuppe;Bajaj, Jitendra;Sharma, Abhishek;Manimaran, Sellappan;Ravanappa, Prashantha Kumar Bommenahalli;Pottekad, Vijayan
    • Advances in Traditional Medicine
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    • v.10 no.4
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    • pp.263-270
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    • 2010
  • Scoparia dulcis L. is widely used in the traditional system of medicine for treating liver ailments. In the present study the terpenoids and terpenoid fractions isolated from 1:1:1 petroleum ether, diethyl ether and methanol (PDM) extract of Scoparia dulcis L. were tested for their in vitro 1, 1-Diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity. Selected samples from the assay were further tested for their in vitro hepatoprotective activity against $CCl_4$ induced hepatotoxicity in freshly isolated rat hepatocytes. In the in vitro antioxidant study, fractions 7, 11, 13, 14, and 15 and PDM extract show the DPPH radical scavenging activity. The phytochemical screening of all these fractions show the presence of terpenoids. In the in vitro hepatoprotective study all these fractions and the PDM extract significantly prevent the $CCl_4$ induced changes in the aspartate aspartate amino transferase, alanine amino transferase and alkaline phosphatase levels (p < 0.05). The above results are comparable with the standard, silymarin. The results of the study indicate that, the PDM extract of Scoparia dulcis L. possesses potential hepatoprotective activity and this may be attributed to its free radical scavenging potential, which in turn may be attributed to the presence of terpenoids.

Clinical outcomes of preimplantation genetic testing for aneuploidy in high-risk patients: A retrospective cohort study

  • Jun Woo Kim;So Young Lee;Chang Young Hur;Jin Ho Lim;Choon Keun Park
    • Clinical and Experimental Reproductive Medicine
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    • v.51 no.1
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    • pp.75-84
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    • 2024
  • Objective: The purpose of this study was to evaluate the impact of preimplantation genetic testing for aneuploidy (PGT-A) on clinical outcomes among high-risk patients. Methods: This retrospective study involved 1,368 patients and the same number of cycles, including 520 cycles with PGT-A and 848 cycles without PGT-A. The study participants comprised women of advanced maternal age (AMA) and those affected by recurrent implantation failure (RIF), recurrent pregnancy loss (RPL), or severe male factor infertility (SMF). Results: PGT-A was associated with significant improvements in the implantation rate (IR) and the ongoing pregnancy rate/live birth rate (OPR/LBR) per embryo transfer cycle in the AMA (39.3% vs. 16.2% [p<0.001] and 42.0% vs. 21.8% [p<0.001], respectively), RIF (41.7% vs. 22.0% [p<0.001] and 47.0% vs. 28.6% [p<0.001], respectively), and RPL (45.6% vs. 19.5% [p<0.001] and 49.1% vs. 24.2% [p<0.001], respectively) groups, as well as the IR in the SMF group (43.3% vs. 26.5%, p=0.011). Additionally, PGT-A was associated with lower overall incidence rates of early pregnancy loss in the AMA (16.7% vs. 34.3%, p=0.001) and RPL (16.7% vs. 50.0%, p<0.001) groups. However, the OPR/LBR per total cycle across all PGT-A groups did not significantly exceed that for the non-PGT-A groups. Conclusion: PGT-A demonstrated beneficial effects in high-risk patients. However, our findings indicate that these benefits are more pronounced in carefully selected candidates than in the entire high-risk patient population.

Comparison of in vitro ruminal fermentation incubated with different levels of Korean corn grains with total mixed ration as a basal

  • Hamid, Muhammad Mahboob Ali;Park, Ha Young;Choi, Chang Weon
    • Korean Journal of Agricultural Science
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    • v.45 no.3
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    • pp.419-427
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    • 2018
  • The present study was conducted to investigate the effect of different levels of Korean corn grain on in vitro ruminal fermentation with total mixed ration (TMR) as a basal feed. Three ruminal cannulated Holstein steers (Body Weight $479{\pm}33.0kg$) were used as rumen fluid donors. Treatments for in vitro fermentation were TMR only (control, 3.0 g), TMR substituted partially with high level (HC, TMR 1.5 and corn 1.5 g), and with low level of Korean corn grain (LC, TMR 2.25 and corn 0.75 g), respectively. To measure in vitro ruminal pH, gas production, ammonia N and volatile fatty acids (VFA), the in vitro fermentation incubation was triplicated at $39^{\circ}C$, 120 rpm for 0, 1, 3, 6, 12, 24 and 48 h, respectively. Mean ruminal pH was significantly lower (p < 0.05) for HC than control. Changes in rumen pH was rather similar between the groups till 6 h after incubation, but the lowest pH for HC (pH 5.10) appeared at 48 h compared with control and LC. Total gas production was tended (p < 0.09) to be higher and ammonia N was significantly lower (p < 0.05) for HC than control and LC. Total VFA was higher (p < 0.05) for HC and LC than control but no differences appeared between HC and LC. Overall, the present data indicate that feeding different levels of Korean domestic corn grain may lead to high and sustainable starch degradation in the rumen.

Improved Preimplantation Development of Cloned Porcine Embryos through Supplementation of Histone Deacetylase Inhibitor MS-275

  • Fang, Xun;Qamar, Ahmad Yar;Shin, Sang Tae;Cho, Jongki
    • Journal of Veterinary Clinics
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    • v.36 no.5
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    • pp.253-258
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    • 2019
  • The objective of this study was to analyse the effects of MS-275 (Class I and II histone deacetylase inhibitor) supplementation on the development of porcine in-vitro somatic nuclear transfer embryo production. During in-vitro development, early embryos were exposed to different concentrations of MS-275 (0, $5{\mu}M$, $10{\mu}M$, and $20{\mu}M$). In in-vitro culture supplemented group, the blastocyst development rate was significantly enhanced by $10{\mu}M$ concentration than other groups (24.0% vs. 19.3%, 21.8%, 11.5%; P < 0.05). Additionally, the 6 h supplementation group, significantly improved the blastocysts production than 24 h, 48 h and control groups (26.1% vs. 17.0%, 15.2%, 2.8%; P < 0.05). Following supplementation with optimal concentrations and time ($10{\mu}M$-6 h group), the blastocyst production was significantly higher than control (25.7% vs 15.8%; P < 0.05). The optimal concentrations of MS-275 significantly enhanced the percentages of ICM:TE than control (43.6% vs. 38.4%; P < 0.05) accompanied with significantly higher expression levels of reprogramming related genes (POU5F1, Naong, and SOX2). In conclusion, the optimal concentrations of $10{\mu}M$ MS-275 and 6 h supplementation during in-vitro culture can significantly improve the quality of porcine in-vitro somatic nuclear transfer embryos through histone acetylation and epigenetic modification. Increasing the efficiency of clonal animal production will greatly promote the development of animal disease models and xenotransplantation.