• Title/Summary/Keyword: co-culture system

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Enhanced Neurite Outgrowth of Dorsal Root Ganglion Sensory Neurons after Sibjeondaebo-tang Treatment

  • Kwon, Ku-Birm;NamGung, Uk
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.24 no.4
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    • pp.681-687
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    • 2010
  • Sibjeondaebo-tang (SJDBT) is an oriental medicinal prescription for the treatments of diverse symptoms including neurological disorders. In order to investigate its potential role for neural regulation following nerve injury, neurite outgrowth of dorsal root ganglion (DRG) neurons in culture was investigated. In DRG neurons which were preconditioned by sciatic nerve injury, neurite outgrowth was enhanced by SJDBT treatment. When preconditioned DRG neurons were co-cultured with astrocytes prepared from injured spinal cord tissue, neurite outgrowth was similarly facilitated by SJDBT. Astrocytes in co-culture showed more intense signals of vimentin protein by SJDBT compared to saline control. Sukjihwang (SJH), a conventional herbal component of SJDBT prescription, did not induce any significant changes in neurite extension of DRG neurons compared to control cells. These data suggest that SJDBT may be the therapeutic agent for nervous system disorders related to nerve damage.

Insulin enhances neurite extension and myelination of diabetic neuropathy neurons

  • Pham, Vuong M.;Thakor, Nitish
    • The Korean Journal of Pain
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    • v.35 no.2
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    • pp.160-172
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    • 2022
  • Background: The authors established an in vitro model of diabetic neuropathy based on the culture system of primary neurons and Schwann cells (SCs) to mimic similar symptoms observed in in vivo models of this complication, such as impaired neurite extension and impaired myelination. The model was then utilized to investigate the effects of insulin on enhancing neurite extension and myelination of diabetic neurons. Methods: SCs and primary neurons were cultured under conditions mimicking hyperglycemia prepared by adding glucose to the basal culture medium. In a single culture, the proliferation and maturation of SCs and the neurite extension of neurons were evaluated. In a co-culture, the percentage of myelination of diabetic neurons was investigated. Insulin at different concentrations was supplemented to culture media to examine its effects on neurite extension and myelination. Results: The cells showed similar symptoms observed in in vivo models of this complication. In a single culture, hyperglycemia attenuated the proliferation and maturation of SCs, induced apoptosis, and impaired neurite extension of both sensory and motor neurons. In a co-culture of SCs and neurons, the percentage of myelinated neurites in the hyperglycemia-treated group was significantly lower than that in the control group. This impaired neurite extension and myelination was reversed by the introduction of insulin to the hyperglycemic culture media. Conclusions: Insulin may be a potential candidate for improving diabetic neuropathy. Insulin can function as a neurotrophic factor to support both neurons and SCs. Further research is needed to discover the potential of insulin in improving diabetic neuropathy.

Effect of Semen Sources and Culture System on Efficiency of IVP Embryo Production and Cryopreservation (정액종류 및 배양조건에 따른 체외수정란의 생산 및 동결보존의 효율에 미치는 영향)

  • 공일근;이상인
    • Journal of Embryo Transfer
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    • v.14 no.1
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    • pp.31-37
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    • 1999
  • The objective of this study was to optimize the selection of sperm sources, optimal culture systems and vitrification method depends on sperm sources. The oocytes were inseminated with either KPN 105, 114, 191, SNU 101, 102, 103 or epididymis and then embryos inseminated were cultured in oviductal cell co-culture or HECM-6 as defined me dium. The blastocysts produced were pooled according to sperm sources as KPN, SNU or epididymis and then vitrified by OPP vitrification method. The results obtained were as follows: 1. The cleavage(86.2 or 84.7%) and development rates to blastocyst (30.6 or 32.0%) were not significantly different between oviductal cell co-culture or HECM-6 culture systems(P<0.05). 2. To determine the optimal sperm sources for using IVF in this system, cleavage rates in KPN 191 and SNU 101 (74.2, 55.8%) were significantly lower rather than those in KPN 105, 114, SNU 102, 103 or epididymis (86.7, 85.1, 89.8, 85.5 or 81.2%), but development rates to blastocyst in KPN 114, SNU 103 or epididymis sperm (30.0, 33.0 or 28.6%) were significantly higher rater than those in KPN 105, 191, SNU 101, 102(21.4, 15.4, 14.9 or 25.4%), respectively (P<0.05). 3. The blastocysts produced were pooled according to sperm sources as KPN, SNU or epididymis and then vitrified by OPP vitrification method. The survival rates were not significantly different among sperm sources (89.6%: 43/48 ; 90.1%: 46/51 ; 83.3% : 20/24). These results obtained indicate that the defined medium, HECM-6, could be use to produce of IVP bovine embryos. Since the frozen semen must be required to maintain of unvariation data in IVP embryo production system, KPN 114 and SNU 103 produced in our laboratory were useful for this purpose. The blastocysts produced by different sperm sources as KPN, SNU or epididymis were vitrified by OPP vitrification method and survived very high rates. The OPP vitrification method could be susceptibility to use of IVP bovine blastocyst embryos.

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Enhancement of Cultivation Efficiency of Bifidobacterium longum Using Calcium Carbonate Buffer System (Calcium Carbonate Buffer System을 이용한 Bifidobacterium longum의 배양 효율 증대에 관한 연구)

  • Lee, Ki-Yong;Hwang, In-Bum;Heo, Tae-Ryeon
    • Korean Journal of Food Science and Technology
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    • v.29 no.1
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    • pp.126-132
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    • 1997
  • Calcium carbonate ($CaCO_3$) immobilized with alginate was studied as buffer system to enhance the cultivation efficiency of Bifidobacterium longum (ATCC 15707) which is inhibited at low pH. To test the bufferring effect of the immobilized $CaCO_3$ beads, pH value in each modified trypticase-proteose peptone-yeast (TPY) broth which is adjusted to pH 4.0 with acetic acid, lactic acid and complex solution of acetic and lactic acid, 3:2 (M:M) was tested by concentration of $CaCO_3$ bead and reaction time. The bufferring effect of $CaCO_3$ bead became higher with increasing the amount of $CaCO_3$ bead in the acidic solution. The growth rate of bifidobacteria and bufferring effect were examined in relation to the amount of $CaCO_3$ bead and concentration of glucose in the modified TPY media. The growth rate of bifidobacteria and bufferring effect were increased with increasing the amount of $CaCO_3$ bead and concentration of glucose. Also, the exponential time of bifidobacteria became longer with increasing the amount of $CaCO_3$ bead and concentration of glucose in the modified TPY media. When we observed the growth rate of bifidobacteria by the method of pH-controlled culture and $CaCO_3$ buffer system, the $CaCO_3$ buffer system was more effective than that of pH-controlled culture. Therefore, this $CaCO_3$ buffer system may be useful as a method to enhance of the cultivation efficiency of bifidobacteria.

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Secretory Production of Biologically Active Human Thrombopoietin by Baculovirus Expression System

  • Koh, Yeo-Wook;Lim, Seung-Wook;Park, Seung-Kook;Park, Myung-Hwan;Na, Doe-Sun;Yang, Jai-Myung
    • BMB Reports
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    • v.31 no.5
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    • pp.453-458
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    • 1998
  • Human thrombopoietin (hTPO) was expressed to high levels in insect cells using the baculovirus expression system. Full-length hTPO cDNA containing a native signal peptide sequence was amplified by PCR from a human fetal liver cDNA library and cloned into the Autographa californica nuclear polyhedrosis virus (AcNPV) expression vector. Immunoblot analysis with antiserum against hTPO indicated that an approximately 55 kDa protein was produced in recombinant AcNPV infected insect cells. Recombinant hTPO was produced 4-fold higher in Trichoplusia ni (Tn5) cells than in Spodoptera frugiperda (Sf9) cells. with most of the hTPO produced in Tn5 cells secreted into the culture medium. Addition of tunicamycin in the culture medium resulted in the reduction of the size of hTPO to 35-38 kDa, and most of the protein remained within the cell. These results suggest that N-glycosylation of hTPO is required for the secretion of the protein into the culture medium in insect cells. hTPO produced in insect cells induced proliferation and maturation of megakaryocyte progenitors, indicating that it is in a biologically active form.

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Ca-Alginate에 고정화된 Calcium Carbonate를 완충제로 사용한 Bifidobacterium longum의 배양 증대와 저장 안정성

  • Lee, Gi-Yong;Yu, Won-Gyu;Kim, Ji-Yeon;Heo, Tae-Ryeon
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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    • pp.203-206
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    • 2000
  • Calcium carbonate $(CaCO_3)$ bead immobilized with alginate were developed as buffer system to enhance the cultivation efficiency of bifidobacteria. When Bifidobacteriuim longum KCTC 3128 and HLC 3742 were independently cultivated in 2.5-liter fermenter buffered the $CaCO_3$ bead, NaOH, $Na_2CO_3$, and $NH_4OH$. The proliferation of bifidobacteria and their storage stability were higher in culture broth buffered $CaCO_3$ beads than in culture broth buffered with NaOH, $Na_2CO_3$, and $NH_4OH$. Therefore, $CaCO_3$ bead may be useful as a buffer to enhance of the cultivation efficiency and viability of bifidobacteria.

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Fermentation Characteristics of Large-scale Coenzyme Q10 Expressing Rhodobacter spharoides in Rumen Simulated Continuous Culture (RSCC) System (Coenzyme Q10 다량 발현 미생물을 이용한 Rumen Simulation Continuous Culture (RSCC) System 반추위 내 미생물 발효 특성에 대한 연구)

  • Bae, G.S.;Yeo, J.M.;Chang, M.B.;Kim, J.N.
    • Journal of Practical Agriculture & Fisheries Research
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    • v.19 no.1
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    • pp.139-151
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    • 2017
  • This study was conducted to confirm the rumen fermentation characteristics of large-scale CoenzymeQ10(CoQ10) producing bacteria R. spharoides in rumen. We conducted in vitro continuous culture test to investigate the characteristics of rumen fermentation with 5% R. spharoides as a direct fed microorganism. A rumen microbial fermentation characteristic has stability at after 12 days for 15 day of experimental period. pH value, NH3-N, microbial protein synthesis, ADF digestibility and NDF digestibility were not shown significantly differences between control and treatment. However, UDP was significantly higher in treatment than control (p<0.05). CoQ10 concentration was 336.0mg/l with 5% R. spharoides. On the other hands, CoQ10 was not detected without R. spharoides. Our study was shown that R. spharoides can produce CoQ10 in rumen environment without harmful effects on rumen fermentation parameter. CoQ10 in rumen may transfer into cow milk through cow metabolism. This strategy might be helpful for producing functional dairy cow milk.

Measuring Safety Culture to Promote Aviation Safety Culture

  • Kim, Dae Ho;Choi, Jeong Yeol
    • Journal of the Ergonomics Society of Korea
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    • v.35 no.2
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    • pp.111-123
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    • 2016
  • Objective: The objective of this research is to study preceding literature on safety culture surveying tools and indicators used in aviation organizations to help the further understanding of aviation safety culture by presenting Korea-Safety Culture Survey Indicator (K-SCSI) as a relevant case. Background: The aviation field puts a great deal of effort in preventive safety management through the application of Safety Management System (SMS), which was co-developed by international aviation organizations such as ICAO and FAA. To successfully operate safety management system, safety culture factors such as the organization member's level of consciousness, attitude and faith regarding safety must be put together. However, the aviation field currently lacks programs to promote safety culture and the exact understanding of some safety culture concepts. Method: This research inquired into the definition of safety culture in the aviation field and the surveying tools used to measure it. It then described the development and application process of the Korea-Safety Culture Survey Indicator (K-SCSI) mainly focusing on case studies. Results: In this research are presented safety culture promoting programs that can be applied to subordinate indicators of K-SCSI such as organization commitment, management involvement, rationality of reward system, employee empowerment and reporting system. Conclusion: For a mature safety culture to settle successfully, it is essential that safety culture survey indicators are developed and applied in a way that fits the organization's features. Also, behavior measuring indicators are required to develop a more objective indicator and thus must be standardized. Application: Cases that deal with the development and application of safety culture measuring tools within the aviation field can be studied and applied in other domains to spread safety culture.

Effects of Heating on Hydroxyl Radical-Generated Toxicity in Mouse Forebrain Tissue Culture

  • Lee, Jeong-Chae;Lim, Kye-Taek
    • Toxicological Research
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    • v.14 no.3
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    • pp.301-306
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    • 1998
  • This experiment was carrid out to know the effects of heating and serum on hydroxyl radicals in embryonic mouse forebrain (cerebrum) culture. The heating to mouse embryonic cerebrum cells in culture was done in a water bath at 43${\circ}C$ for 60min. After that, two supernatants were prepared at 20 hrs and 48 hrs respectively after heat treatment to the brain cells. To find out the heating effects on neuron cells, mouse cerebrum cells (13 embryonic day) were cultured in hydroxyl radical generation system composed of 20mU/ml glucose oxidase (GO system), using condition of normal culture media (MEM, 5% serum, 5% $CO_2$or supernatant prepared after heating at 43${\circ}C$ for 60 min in a water bath. Supernatant prepared at 20 hrs after heat treatment had a greater protective effects against hydroxyl radical than supernatant prepared at 48 hrs after heat treatment . Otherwise, the protective effect of serum against hydroxyl radicals in the cultured brain cells is higher than that in the heat treatment. These results indicated that serum in culture media reduced cytotoxicity of hydroxyl radicals in mouse forebrain culture, also that heat treatment showed the protective effects against hydroxyl radicals generated with 20mU/ml GO system in mouse forebrain culture.

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