• Title/Summary/Keyword: Suspended cell

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A demonstration of the H3 trimethylation ChIP-seq analysis of galline follicular mesenchymal cells and male germ cells

  • Chokeshaiusaha, Kaj;Puthier, Denis;Nguyen, Catherine;Sananmuang, Thanida
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.6
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    • pp.791-797
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    • 2018
  • Objective: Trimethylation of histone 3 (H3) at 4th lysine N-termini (H3K4me3) in gene promoter region was the universal marker of active genes specific to cell lineage. On the contrary, coexistence of trimethylation at 27th lysine (H3K27me3) in the same loci-the bivalent H3K4m3/H3K27me3 was known to suspend the gene transcription in germ cells, and could also be inherited to the developed stem cell. In galline species, throughout example of H3K4m3 and H3K27me3 ChIP-seq analysis was still not provided. We therefore designed and demonstrated such procedures using ChIP-seq and mRNA-seq data of chicken follicular mesenchymal cells and male germ cells. Methods: Analytical workflow was designed and provided in this study. ChIP-seq and RNA-seq datasets of follicular mesenchymal cells and male germ cells were acquired and properly preprocessed. Peak calling by Model-based analysis of ChIP-seq 2 was performed to identify H3K4m3 or H3K27me3 enriched regions ($Fold-change{\geq}2$, $FDR{\leq}0.01$) in gene promoter regions. Integrative genomics viewer was utilized for cellular retinoic acid binding protein 1 (CRABP1), growth differentiation factor 10 (GDF10), and gremlin 1 (GREM1) gene explorations. Results: The acquired results indicated that follicular mesenchymal cells and germ cells shared several unique gene promoter regions enriched with H3K4me3 (5,704 peaks) and also unique regions of bivalent H3K4m3/H3K27me3 shared between all cell types and germ cells (1,909 peaks). Subsequent observation of follicular mesenchyme-specific genes-CRABP1, GDF10, and GREM1 correctly revealed vigorous transcriptions of these genes in follicular mesenchymal cells. As expected, bivalent H3K4m3/H3K27me3 pattern was manifested in gene promoter regions of germ cells, and thus suspended their transcriptions. Conclusion: According the results, an example of chicken H3K4m3/H3K27me3 ChIP-seq data analysis was successfully demonstrated in this study. Hopefully, the provided methodology should hereby be useful for galline ChIP-seq data analysis in the future.

Studies on Mass Production of Intracellularly-Produced Secondary Metabolite, Cyclosporin A by Use of Immobilized Fungal Cells in Stirred-Tank Immobilized Perfusion Reactor System(IPRS) (교반식 perfusion 생물반응기(IPRS)에서 고밀도 고정상 곰팡이 세포를 이용한 세포내 축적 이차대사산물인 Cyclosporin A 대량생산에 관한 연구)

  • 전계택;이태호장용근
    • KSBB Journal
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    • v.11 no.1
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    • pp.22-29
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    • 1996
  • Immobilized bioprocess was carried out for continuous production of cyclosporin A (CyA) produced intracellularly as a secondary metabolite by a filamentous fungus, Tolypocladium inflatum. Immobilization procedure for entrapping conidiospores of the producer was significantly simplified by use of a modified immobilization technique. A newly-designed immobilized perfusion reactor system (IPRS) showed good process benefits as demonstrated by the role of the high density immobilized cells as an efficient biomass generator, continuously supplying highly active CyA-producing free cells (1.0g/$\ell$/hr) even at very high dilution rate ($0.1hr^{-1}$). IPRS bioprocess was possible since efficient decantor system developed in our laboratory separated the sloughed-off free cells from the immobilized biomass effectively, thus overcoming wash-out phenomenon frequently encountered in continuous free cell cultures. Furthermore the released-free cells remaining in the bulk solution did not appear to cause substrate mass transfer limitation which was often experienced in suspended mycelial fungal cell fermentations. The primary reason for this was that the suspension broth of the IPRS mainly consisted of roundshaped short mycelial fragments and conidiospores, still remaining Newtonian even at high cell density. In parallel with IPRS bioprocess development, other key factors to be considered necessarily for significant increase in CyA productivity would be strain improvement and medium optimization for the immobilized cells.

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Somatic Embryogenesis and Plant Regeneration in Embryogenic Cell Suspension Cultures of Hovenia dulcis Thunb (헛개나무의 현탁배양세포로부터 체세포배발생과 식물체 재생)

  • Li, Cheng-Hao;Zhao, Bo;Kim, Na-Young;Kim, Myong-Jo;Cho, Dong-Ha;Lee, Dong-Wook;Lee, Jae-Geun;Lim, Jung-Dae;Yu, Chang-Yeon
    • Korean Journal of Medicinal Crop Science
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    • v.14 no.4
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    • pp.255-260
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    • 2006
  • Culture conditions for high frequency plant regeneration via somatic embryogenesis from embryogenic cell suspension cultures of Hovenia dulcis are described. Germinated somatic embryos were selected for induction of secondary embryogenesis. Friable embryogenic cells were induced directly from somatic embryos when transfer to 1/3 MS solid or liquid medium lacking plant growth regulators. The temperature strongly effected on induction of secondary embryognesis than other conditions in culture. All somatic embryos produced friable embryogenic cell clumps within 10 days when germinated somatic embryos cultured in 1/3 MS medium at $30^{\circ}C$ in suspension culture. No somatic embryos formed from embryogenic cell suspension cultures at $18^{\circ}C$. Numerous somatic embryos were induced and subsequently developed uniformly into germination stage from suspended cell clumps after 4 weeks of culture on $18^{\circ}C$. Plantlets conversion were observed on $18^{\circ}C$ when germinated somatic embryos were transferred to 1/3 MS solid medium without plant growth regulators or supplemented with 0.1-0.5 mg/l benzyladenine.

The Effect of the Aging of Red Blood Cells on Rheological Properties and Hemolysis

  • Tomioka, Jun;Motokubo, Kazuhiro;Watanabe, Hisayoshi
    • Proceedings of the Korean Society of Tribologists and Lubrication Engineers Conference
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    • 2002.10b
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    • pp.371-372
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    • 2002
  • It is well known that red blood cells (RBCs) are suffered from chronic stresses in systemic circulation. The objective of this study is to clarify the effect of the aging of RBCs on rheological properties and hemolysis. Initially, RBCs age fractionation was performed by using a high-speed centrifugation (15[min] at 1500[G]), then young and aged RBCs were suspended in plasma to adjust the hematocrit level of 40[%]. After this pretreatment, the viscosity was measured by using a capillary type and a cone-plate type viscometers, respectively, and the hemolysis test was carried out by a seesaw type shaker. Results from these experiments showed that the viscosity of the aged RBCs measured by the capillary viscometer was increased by 10[%] as compared with that of the young RBCs. Under the condition of all shear zones, the viscosity of the aged RBCs was increased in case of using the cone-plate type viscometer. And the hemolytic level was increased twice as the aging. The data obtained in this study indicated that the ability of aggregation of RBCs was increased and the deformability of RBCs membrane got lower with the aging. Furthermore, it was exhibited that the fragility of RBCs ’ membrane was increased with the aging.

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In vitro functional assenssment of bioartificial liver system using immobilized porcine hepatocyte spheroids

  • Lee, Ji-Hyun;Lee, Doo-Hoon;Yoon, Hee-Hoon;Jung, Doo-Hee;Park, Jung-Keug;Kim, Sung-Koo;Lee, Kwang-Woong;Lee, Suk-Koo
    • 한국생물공학회:학술대회논문집
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    • 2003.10a
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    • pp.305-306
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    • 2003
  • To treat fulminant hepatic failure (FHF) patients, various extracorporeal bioartificial liver (BAL) systems have been developed. Several requirements should be met for the development of BAL systems: hepatocytes should be cultured in a sufficiently high density; their metabolic functions should be of a sufficiently high level and duration; and the BAL systems module should permit scaling-up and aseptic handling. Several investigators have found that freshly isolated primary hepatocytes can be cultured into three dimensional, tightly packed, freely suspended, multicellular aggregates, or spheroids. These specialized cell structures exhibited enhanced liver specific functions and a prolonged differentiated state compared to cells maintained in a monolayer culture. Cells in spheroids appear to mimic the morphology and ultrastructure of the in vivo liver lobule. The ability of hepatocytes to organize into three-dimensional structures was hypothesized to contribute to their enhanced liver-specific activities. In this study, the ammonia removal rate and urea secretion rate of pig hepatocytes spheroids encapsulated in Ca-alginate bead were determined. A packed-bed bioreactor with encapsulated pig hepatocytes was devised as BAL support system. The efficacy of the system was evaluated in vitro.

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The Therapeutic Effect of Natural Honeybee (Apis mellifera) Venom in Adjuvant-induced Arthritic Rat (관절염 유발 랫드에 대한 생봉독의 치료 효과)

  • 강성수;최석화;조성구
    • Journal of Veterinary Clinics
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    • v.16 no.1
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    • pp.155-162
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    • 1999
  • This study was performed to assess that clinco-therapeutic effect of natural Italian honeybee (Apis mellifera) venom in adjuvant-induced arthritic rat. Ninety Sprague- Dawley rats of male were injected with complete Freund's adjuvant (CFA). Adjuvant arthritis was produced by a single subcutaneous injection of 1 mg Mycobacterium butyricum suspended in 0.1 ml paraffin oil into the right hindpaw. Righting reflex was uniformly lost and considered to be the point of arthritis development on day 14 after CFA injection. Experimental groups were divided into three groups. When arthritis was developed in the rat hind-paw, tested groups were administrated with prednisolone (10 mg/kg, p.o) and honeybee venom (one bee, s.c) at an interval of two days. Control group was subcutaneously injected with 0.1 ml of physiological saline solution in the rat at an interval of two days. Clinical findings, hematological values and histopathological findings were observed during or after the drugs administration. In tested groups, the development of inflammatory edema and polyarthritis on day 14 after treatment was suppressed. No significant differences of hindpaw edema volume and lameness score between prednisolone and honeybee venom groups were observed during or after therapeutic drugs treatment. WBC counts of prednisolone and honeybee venom treatment groups as compared with the control group were getting remarkably decreased during or after the therapeutic drugs administration(p<0.01). Erosions of articular cartilage and inflammatory cell infiltrations during or after the therapeutic drugs treatment was effectively suppressed in natural honey venom.

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Flora of drift plastics: a new red algal genus, Tsunamia transpacifica(Stylonematophyceae) from Japanese tsunami debris in the northeast Pacific Ocean

  • West, John A.;Hansen, Gayle I.;Hanyuda, Takeaki;Zuccarello, Giuseppe C.
    • ALGAE
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    • v.31 no.4
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    • pp.289-301
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    • 2016
  • Floating debris provides substrates for dispersal of organisms by ocean currents, including algae that thrive on plastics. The 2011 earthquake and tsunami in Tohuku, Japan resulted in large amounts of debris carried by the North Pacific Current to North America from 2012 to 2016. In 2015-2016, the plastics in the debris bore a complex biota including pink algal crusts. One sample (JAW4874) was isolated into culture and a three-gene phylogeny (psbA, rbcL, and SSU) indicated it was an unknown member of the red algal class Stylonematophyceae. It is a small pulvinate crust of radiating, branched, uniseriate filaments with cells containing a single centrally suspended nucleus and a single purple to pink, multi-lobed, parietal plastid lacking a pyrenoid. Cells can be released as spores that attach and germinate to form straight filaments by transverse apical cell divisions, and subsequent longitudinal and oblique intercalary divisions produce masses of lateral branches. This alga is named Tsunamia transpacifica gen. nov. et sp. nov. Sequencing of additional samples of red algal crusts on plastics revealed another undescribed Stylonematophycean species, suggesting that these algae may be frequent on drift oceanic plastics.

Water Quality Monitoring for Hazard Analysis in Aquaculture Farm of Rainbow Trout (송어양식장의 위해요소 관리를 위한 수질 모니터링)

  • Kim, Young-Mog;Lee, Myung-Suk;Chung, Yong-Hyun
    • Journal of Fisheries and Marine Sciences Education
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    • v.25 no.4
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    • pp.819-827
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    • 2013
  • Water quality has been considered to be one of sanitation standard operating procedures (SSOP) for hazard analysis critical control points (HACCP) application in aquaculture farms. This study was conducted to evaluate a hazard caused by water used in aquaculture farm of rainbow trout. The water quality was analyzed to investigate both physiochemical and bacteriological level in water samples collected from aquaculture farm of rainbow trout, Oncorhynchus mykiss. No significant difference were observed on water temperature and pH from season to season. However, the levels of dissolved oxygen were decreased as the outside temperature was increased, even if the levels were adequate for aquaculture. Also, other physiochemical analysis including biochemical oxygen demand (BOD), chemical oxygen demand (COD) and suspended solid (SS) revealed that the waters for aquaculture analyzed in this study was suitable for rainbow trout aquaculture. The bacterial analyses were also revealed that the waters for aquaculture were met to both coliform group (<18 MPN/100mL) and viable cell count (<100 CFU/mL). However, some of waste waters from aquaculture farms showed higher levels of BOD and COD than those of waste water standard (<2 ppm), suggesting that regular cleaning of fish tank and precipitation tank is needed.

Recycling of Wastepaper(V): -Calcium Hardness Control of Process Water for Zero-Discharge System- (고지재상연구 (제5보) -공정수 폐쇄화를 위한 칼슘경도 조절-)

  • 지경락;류정용;신종호;송봉근;오세균
    • Journal of Korea Technical Association of The Pulp and Paper Industry
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    • v.31 no.2
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    • pp.15-24
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    • 1999
  • A new technique for recycling of white water was developed in order to reduce the calcium hardness in a closed OCC recycling system. Calcium ions present in the white water were precipitated as calcium carbonate by reacting with sodium carbonate, and the precipitated $CaCO_3$ was removed from the system using a flotation fractionation method, which has been commonly used in deinking process. In the flotation stage, a mixed gas of $CO_2$-air was purged into the flotation cell because the pH of $Na_2CO_3$-treated white water was reduced to neutral by $CO_2$ gas. Since $CaCO_3$ precipitate tends to stick onto fine fiber surface and then selectively removed from the white water, a proper amount of suspended solid in white water acts as an important factor for deciding the removal efficiency. By the application of $Na_2CO_3$ addition-$CO_2$ flotation to the short circulated white water, the calcium hardness was significantly reduced by 87% and more. Removal of calcium ions with fine fibers led to a drainage improvement, reduction of fresh water consumption, and enhanced efficiency of wet-end chemicals.

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Evaluation on Light Scattering Behavior of a Pulverized Coal Suspension (슬러리내 석탄입자의 광산란 특성 평가)

  • Hwang, Munkyeong;Nam, Hyunsoo;Kim, Kyubo;Song, Juhun
    • Journal of Hydrogen and New Energy
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    • v.24 no.5
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    • pp.451-460
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    • 2013
  • In a direct coal fuel cell (DCFC) system, it is essential to identify volume fraction of coal suspended in electrolyte melt in order to control its dispersion and fluidity. This requirement is compelling especially at anode channel where hot slurry is likely to flow at low velocity. In this study, light scattering techniques were employed to measure the volume fraction for a pulverized coal suspension with relatively high absorption coefficient. The particle size, scattering angle, and volume fraction were varied to evaluate their effects on the scattering behavior as well as scattering regime. The larger coal size and smaller forward scattering angle could provide a shift to more favorable scattering regime, i.e., independent scattering, where interferences of light scattering from one particle with others are suppressed.