• Title/Summary/Keyword: prolonged culture

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Effects of Oocyte Maturational Age and Activation Conditions on the Development of Porcine Parthenogenetic Embryos

  • Kwon, Dae-Jin;Park, Joo-Hee;Park, Choon-Keun;Yang, Boo-Keun;Cheong, Hee-Tae
    • Reproductive and Developmental Biology
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    • v.31 no.2
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    • pp.77-82
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    • 2007
  • This study was conducted to investigate the effects of oocyte maturational age and activation condition on in vitro development of porcine parthenogenetic embryos (parthenotes). Porcine follicular oocytes were matured in vitro for 30 to 44 hr. Maturation rate was examined during in vitro maturation (IVM) every 2 hr interval. The cdc2 kinase activity was measured at 36 and 44 hr of IVM. Some oocytes were activated at 36 or 44 hr of IVM by three different conditions; 1) single electric stimulation (1.5 kV/cm for $30{\mu}sec$; ES), 2) double electric stimulations (1.5 kV/cm for $30{\mu}sec$, followed by 1.0 kV/cm for $50{\mu}sec$ after 1 hr; ES+ES) or 3) ES+ES followed by culture in 6-dimethlyaminopurine (6-DMAP) for 4 hr (ES+ES+D), and cultured for 6-7 days. Maturation rate was significantly increased as culture period was increased to 36 hr (66.9%, p<0.05), and then gradually increased to 87.1% at 44 hr of IVM. The cdc2 kinase activity was decreased (p<0.05) with culture period prolonged from 36 hr to 44 hr. Lower blastocyst formation rate (4.3%, p<0.05) were obtained by ES in 36 hr-matured oocytes compared to other treatments (16.5 and 20.5%) in the same age and the same treatment in 44 hr-matured oocytes (15.0%). High blastocyst formation rate (23.6%) was obtained by ES+ES+D in 44 hr-matured oocytes (p<0.05). These results demonstrate that porcine oocyte activation and in vitro development of parthenotes can be affected by interactions between oocyte maturational age and activation condition.

Plant Regeneration and Somatic Embryo Formation from Root-Derived Callus of Rice (벼 뿌리조직 유래의 캘러스로부터 체세포배 형성과 식물체 재분화)

  • 손재근;김경민;김종수
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.3
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    • pp.143-148
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    • 1995
  • The competence of callus formation and plant regeneration from root derived callus was higher in japonica cultivars than those of Tongil-type cultivars of rice. A japonica type cultivars Yeongdeogbyeo, showed the highest capacity (13%) for plant regeneration from root calli of 6 cultivars tested. The callus induced from seed and root tissues maintained higher capacity for plant regeneration during 7 passages of subculture on N$_{6}$ solid media at 2-week intervals. The maximum frequency (2 x 10$^{5}$ mL) of round cells and their cell colonies showed about 24 days after suspension culture of root-derived callus in N$_{6}$ medium with lmg/L 2,4-D, 300mg/L casein hydrolysate, 10mM L-proline, 20g/L sucrose and 30g/L sorbitol. The frequency of somatic embryo formation in suspension cultures of root-derived callus increased with prolonged advance of subculture time from 30 to 90 days, but their regenerative capacities decreased.

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Effects of Starvation on Lipid Metabolism and Gluconeogenesis in Yak

  • Yu, Xiaoqiang;Peng, Quanhui;Luo, Xiaolin;An, Tianwu;Guan, Jiuqiang;Wang, Zhisheng
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.11
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    • pp.1593-1600
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    • 2016
  • This research was conducted to investigate the physiological consequences of undernourished yak. Twelve Maiwa yak ($110.3{\pm}5.85kg$) were randomly divided into two groups (baseline and starvation group). The yak of baseline group were slaughtered at day 0, while the other group of yak were kept in shed without feed but allowed free access to water, salt and free movement for 9 days. Blood samples of the starvation group were collected on day 0, 1, 2, 3, 5, 7, 9 and the starved yak were slaughtered after the final blood sample collection. The liver and muscle glycogen of the starvation group decreased (p<0.01), and the lipid content also decreased while the content of moisture and ash increased (p<0.05) both in Longissimus dorsi and liver compared with the baseline group. The plasma insulin and glucose of the starved yak decreased at first and then kept stable but at a relatively lower level during the following days (p<0.01). On the contrary, the non-esterified fatty acids was increased (p<0.01). Beyond our expectation, the ketone bodies of ${\beta}$-hydroxybutyric acid and acetoacetic acid decreased with prolonged starvation (p<0.01). Furthermore, the mRNA expression of lipogenetic enzyme fatty acid synthase and lipoprotein lipase in subcutaneous adipose tissue of starved yak were down-regulated (p<0.01), whereas the mRNA expression of lipolytic enzyme carnitine palmitoyltransferase-1 and hormone sensitive lipase were up-regulated (p<0.01) after 9 days of starvation. The phosphoenolpyruvate carboxykinase and pyruvate carboxylase, responsible for hepatic gluconeogenesis were up-regulated (p<0.01). It was concluded that yak derive energy by gluconeogenesis promotion and fat storage mobilization during starvation but without ketone body accumulation in the plasma.

Control of spermatozoa penetration and polyspermy by cumulus cells in porcine oocytes matured in culture

  • Shin, Myung-Kyun;Chung, Kee-Soo;Chung, Dong-Soo
    • Korean Journal of Veterinary Service
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    • v.26 no.4
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    • pp.329-337
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    • 2003
  • The functional role of the cumulus cells on sperm penetration and polyspermy during in vitro fertilization was examined. The penetration rate was significantly higher(p<0.01) in oocytes with(61%) than without(25%) cumulus cells. No significant differences, however, was observed in polyspermy. When the hyaluronidase was supplemented to the fertilization medium with different concentrations, penetration rates in oocytes with cumulus cells were higher than oocytes without cumulus cells at 0(61 vs 34% ; p<0.05), 0.01(56 vs 35% ; p<0.05), 0.1(66 vs 30% ; p<0.05) and 1.0 mg/$m\ell$(39 vs 27%). On the other hand, the polyspermy rates were lower oocytes without than with cumulus cells, and had a tendency to decrease with high concentrations of hyaluronidase. In another experiment, the penetration and polyspermy rates had a tendency to increase as time of sperm-oocytes culture was prolonged. At 16 and 20hrs after insemination, the penetration rates were significantly higher(p<0.05) in oocytes with(48 and 62% for 16 and 20hrs) than without(25 and 31% for 16 and 20hrs) cumulus cells in medium with hyaluronidase. However, the polyspermy rates were significantly(p<0.05) lower in oocytes without(3 and 16%) than with(37 and 48%) cumulus cells at 16 and 20hrs after insemination. In cumulus-free oocytes inseminated in medium with or without hyaluronidase at different concentrations of cumulus cells, the penetration rates were significantly(p<0.05) higher in medium with than without hyaluronidase at different concentrations of cumulus cells. The proportions of polyspermy were lower in medium without than with hyaluronidase at 0 (10 vs 0%), 10$^2$(25 vs 0%), 10$^4$(24 vs 14%) and 10$\^$6/(29 vs 10% ; p<0.05) cumulus cells/ml. These results suggest the advantage of culture in medium with cumulus cells and denuded oocytes to inhibit polyspermy with no decrease in the penetration rates during the fertilization in vitro in the porcine.

Variations of Agronomic Traits on the Progenies of the Different Callus Origin and Subculture Period in Rice (벼에서 캘러스 유기원와 계대배양 기간에 따른 재분화 계통의 주요 농업형질 변이)

  • Yi, Gi-Hwan;Oh, Byeong-Geun;Yang, Sae-Jun;Kim, Soon-Chul;Nam, Min-Hee
    • Journal of Plant Biotechnology
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    • v.30 no.4
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    • pp.365-368
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    • 2003
  • Variations of agronomic traits were evaluated on the progenies of regenerated rice plants of different callus origin and subculture period. From eighty-eight percent to ninety percents of ovary culture-derived plants (OCP) and anther culture-derived plants (ACP) were not segregated within the lines in major agronomic characters. Compare to ACP and OCP lines, the seed culture derived lines (SCP) showed more segregation (17%) in major agronomic traits among the lines. The most frequent segregating traits were grain fertility (6.1%) in ACP, leaf color (4.2%) in OCP and grain fertility (4.9%) and leaf color (4.9%) in SCP lines. The SCP line showed more variation in culm length, panicle length and heading date than those of OCP and ACP lines. The variation of agronomic traits in SCP lines was tended to increase with prolonged subculture. Culm and panicle length were shorter than those of original cultivar in all three types of tissue culture-derived lines (OCP, ACP and SCP).

Use of a Xanthine-Xanthine Oxidase System on In Vitro Maturation and Fertilization in Pig

  • Sa, S.J.;Park, C.K.;Cheong, H.T.;Yang, B.K.;Kim, C.I.
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.13-13
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    • 2001
  • This study was undertaken to evaluate the effects of catalase using xanthine (X) - xanthine oxidase (XO) system on in vitro maturation and fertilization in pig. When follicular oocytes were cultured in maturation medium with X and/or XO, the maturation rates were not significantly different between in medium with and without catalase despite of different culture periods. However, significantly (P<0.05) higher maturation rates were obrained in culture with X-XO system. The rates of degenerated oocytes were increased with culture periods prolonged, and were significantly (P<0.05) higher in medium without than with catalase at 120 h of culture. On the other hand, the parthenogenetic oocytes were observed with high proportions at 72 h of culture, hut were not different in medium with and without catalase at various times of culture. In another experiment, the frozen-thawed boar spermatozoa treated with X-XO system for in vitro fertilization. The penetration rates were higher in medium with that than without catalase during the in vitro fertilization with, none (P<0.05), XO and X+XO. On the other hand, when sperm were treated with none, X, XO and X+XO, lipid peroxidation were higher in medium without that than with catalase. However, the changes in sperm penetration and lipid peroxidation showed opposite patterns. The sperm suspensions were also treated with X and/or XO for assay of sulfhydryl (-SH) group content. Under the above all conditions, sperm-SH group were higher detected In medium with that than without catalase. The activity of sperm binding to zona pellucida was also evaluated through binding to salt-stored porcine oocytes. In control group, sperm binding to zona pellucida were higher than in medium with X, XO and X+XO groups. No significant differences, however, were observed between medium with and without catalase. In conclusion, the exposure of follicular oocytes and spermatozoa to X-XO system may be caused stimulating in vitro maturation and fertilization in pig. This work was supported by grant No. 2000-1-22200-001-3 from the Basic Research Program of the Korea Science & Engineering Foundation.

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Morphology and Chemical Composition Analysis of Human Cremated Ash by SEM/EDS (SEM/EDS를 이용한 화장 분골의 형태와 성분 분석)

  • Hwang, Kyu-Sung;An, Woo-Hwan;Kim, Jeong-Lae
    • The Journal of the Convergence on Culture Technology
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    • v.1 no.2
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    • pp.65-69
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    • 2015
  • Teeth and bones are very resistance to high temperatures and remain recognizable even after prolonged exposures to heat. The effects of heating and burning on teeth have been studied with the aim of discerning a characteristic signature withstanding high temperature, but there have been few studies about a human cremated ash, especially Korea. We are recognizable by elemental composition and can be detected in human cremated ash samples by Scanning electromicroscopy/Energy dispersive X-ray spectrometry analysis(SEM/EDS), cremated, at $800{\sim}900^{\circ}C$ for 1 hour. In this temperature range, different crystals morphologies(spherical, irregular and hexagonal) are observed in SEM. Calcium(Ca) and oxygen(O) increases steadily after cremation in EDS. We suggest that cremated bone have been provided with calcium oxide(CaO) formation at temperature above $900^{\circ}C$. This study offers basic data to assess the structure and elemental compositions of human ash and to determine if these remain identifiable after exposure to extreme temperatures.

CD30-Mediated Regulation of Cell Adhesion Molecule Expression on Murine T Cells

  • Nam, Sang-Yun
    • IMMUNE NETWORK
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    • v.3 no.1
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    • pp.8-15
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    • 2003
  • Background: CD30 is a member of TNF receptor family and expressed on lymphocytes and other hematopoietic cells following activation as well as Hodgkin and Reed-Sternberg cells in Hodgkin's lymphoma. In this study, CD30-mediated regulation of cell adhesion molecule expression on normal activated mouse T cells was investigated. Methods: Mouse T cells were activated with anti-CD3 antibody for induction of CD30, which was cross-linked by immobilized anti-CD30 antibody. Results: High level of CD30 expression on T cells was observed on day 5, but only little on day 3 even under culture condition resulting in an identical T cell proliferation, indicating that CD30 expression requires a prolonged stimulation up to 5 days. Cross-linking of CD30 alone altered neither proliferation nor apoptosis of normal activated T cells. Instead, CD30 appeared to promote cell adherence to culture substrate, and considerably upregulated ICAM-1 and, to a lesser extent, ICAM-2 expression on activated T cells, whereas CD2 and CD18 (LFA-1) expression was not affected. None of cytokines known as main regulators of ICAM-1 expression on tissue cells (IL 4, $IFN{\gamma}$ and $IFN{\alpha}$) enhanced ICAM-1 expression in the absence of CD30 signals. On the other hand, addition of $NF-{\kappa}B$ inhibitor, PDTC (0.1 mM) completely abrogated the CD30-mediated upregulation of ICAM-1 expression, but not CD2 and ICAM-2 expression. Conclusion: This results support that CD30 upregulates ICAM-1 expression of T cell and such regulation is not mediated by higher cytokine production but $NF-{\kappa}B$ activation. Therefore, CD30 may play important roles in T-T or T-B cell interaction through regulation of ICAM-1, and -2 expression.

Growth Characteristics of Rhoodotorula sp. Y-55 on Ethanol, Acetic acid, and Acetaldehyde Substrates (Ethanol, Acetic acid, Acetaldehyde 기질에서의 Rhoodotorula sp. Y-55의 증식 특징)

  • Yeehn Yeeh;Sang Hae Kim;Woo Hong Joo;Hong Ki Jun;Oh Change Kwon
    • KSBB Journal
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    • v.11 no.3
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    • pp.367-373
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    • 1996
  • The growth characteristics of Rhodotorula sp. Y-55 were examined on minimal medium containing ethanol, acetic acid or acetadehyde as a sole carbon source by batch culture. The increased concentration of substrate reduced overall growth yield and prolonged lag time. The specific growth rate of the yeast was changed, depending upon the initial concentrations of ethanol and acetaldehyde during the exponential period, but was constant on acetic acid without regard to the initial substrate concentrations, giving a value of 0.l07h-1. The highest ${\mu}$ value was obtained on ethanol and acetadehyde substrates and the respective values were 0.270 at 20g/L and 0.041h-1 at 0.2g/L. The maximum overall growth yields were appeared to be 32.6% for ethanol of 10g/L, 25.6% for acetic acid of 20g/L, and 45% for acetaldehyde of 0.2g/L. The respective cellular contents of crude protein and nucleic acids were determined to be 41.5 and 4.9wt% on ethanol and 40.2 and 4.7wt% at the concentration revealing maximal growth yield.

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Expression of Chemokine Receptors Involved in Receptor-Mediated Endocytosis of Bone Marrow-Derived Stromal Stem Cells (골수 유래 기질 줄기세포의 탐식작용 매개성 케모카인 수용체 발현 연구)

  • Jeong, Young-Sin;Byun, Hyang-Min;Shin, Jee-Young;Kim, Jung-Mogg;Chung, Hyung-Min;Oh, Yu-Kyoung
    • Journal of Pharmaceutical Investigation
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    • v.33 no.4
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    • pp.281-286
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    • 2003
  • To design gene deliver systems which can deliver higher amounts of genes into stem cells, we studied the expression of receptors involved in the receptor-mediated endocytosis of bone marrow stromal stem cells. Bone marrow was isolated from ICR mice, and bone marrow stromal stem cells were isolated based on their plastic adherence property. Several culture conditions were screened for effective and continuous culture of marrow stromal stem cells. MesenCult medium was finally used to cultivate marrow stromal stem cells in vitro. As candidate receptors, various chemokine receptors were studied. Both bone marrow cells ad marrow-derived stromal stem cells showed expression of CC chemokine receptors (CCR) and CXC chemokine receptors (CXCR). Marrow stromal stem cells showed higher expression of CCR5 ad CXCR4 chemokine receptors as compared to other types of chemokine receptors. Moreover, though the expression of chemokine receptors generally decreased in most chemokine receptors with the cultivaton of marrow stromal stem cells, CCR5 and CXCR4 chemokine receptors retained the higher level of receptor expressions over prolonged periods. These results suggest that the ligands exhibiting specific binding to CCR5 or CXCR4 might be used to modify gene delivery systems for increased levels of receptor-mediated gene delivery into stromal stem cells.