• Title/Summary/Keyword: co-culture system

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Use of Peristeum as a Source of Endothelial-like Cells (혈관내피유사세포 채취의 원천으로 골막의 활용)

  • Park, Bong-Wook;Kim, Shin-Won;Kim, Uk-Kyu;Hah, Young-Sool;Kim, Jin-Hyun;Kim, Deok-Ryong;Sung, Iel-Young;Cho, Yeong-Cheol;Son, Jang-Ho;Kim, Jong-Ryoul;Byun, June-Ho
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.33 no.5
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    • pp.385-391
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    • 2011
  • Purpose: The periosteum is a well-known source of osteogenic precursor cells for tissue-engineered bone formation. However, cultured endothelial or endothelial-like cells derived from periosteum have not yet been investigated. This study focused on endothelial-like cell culture from the periosteum. Methods: Periosteal tissues were harvested from the mandible during surgical extraction of lower impacted third molars. The tissues were treated with 0.075% type I collagenase in phosphate-buffered saline (PBS) for 1 hr at $37^{\circ}C$ to release cellular fractions. The collagenase was inactivated with an equal volume of DMEM/10% fetal bovine serum (FBS) and the infranatant was centrifuged for 10 min at 2,400 rpm. The cellular pellet was filtered through a $100{\mu}m$ nylon cell strainer, and the filtered cells were centrifuged for 10 min at 2,400 rpm. The resuspended cells were plated into T25 flasks and cultured in endothelial cell basal medium (EBM)-2. Results: Among the hematopoietic markers, CD146 was more highly expressed than CD31 and CD34. The periosteal-derived cells also expressed CD90 and CD166, mesenchymal stem cell markers. Considering that the expression of CD146 was constant and that the expression of CD90 was lower at passage 5, respectively, the CD146 positive cells in passage 5 were isolated using the magnetic cell sorting (MACS) system. These CD146 sorted, periosteal-derived cells formed tube-like structures on Matrigel. The uptake of acetylated, low-density lipoprotein, labeled with 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI-Ac-LDL) was also examined in these cells. Conclusion: These results suggest that the CD146-sorted positive cells can be referred to as periosteal-derived CD146 positive endothelial-like cells. In particular, when a co-culture system with endothelial and osteoblastic cells in a three-dimensional scaffold is used, the use of periosteum as a single cell source would be strongly beneficial for bone tissue engineering.

A Study on Evaluation System of Track Support Stiffness for Concrete Tracks (콘크리트궤도의 궤도지지강성 평가시스템에 관한 연구)

  • Choi, Jung-Youl;Kim, Man-Hwa;Kim, Hyun-Soo;Chung, Jee-Seung
    • The Journal of the Convergence on Culture Technology
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    • v.6 no.2
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    • pp.535-541
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    • 2020
  • A conventional elastic material replacement and performance evaluation are very complicated and time-consuming, and it is difficult to know when to replace the elastic material in advance. By comparing with the product limit and the functional limit, the necessity of elastic material replacement and the improvement of track support stiffness according to replacement can be immediately demonstrated based on experimental data. Using an evaluation system of track support stiffness, the performance evaluation data for elastic materials obtained through field tests using software for track support stiffness is integrated and managed on the administrator's computer. Therefore, the replacement plan is established and maintenance history is managed by identifying the replacement time and location of elastic materials. It is possible to evaluate the performance and condition of the elastic material at the various points during the working time of the track inspection and the track performance (track support stiffness) and durability of the elastic material (aging level, spring stiffness variation rate, etc.) at the operation condition. The elastic material could be replaced timely, and the deterioration of the elastic material can be continuously monitored.

Influence of Ammonium Phosphate on Mycelial Morphology during Submerged Cultivation of Ganoderma lucidum (영지의 액체배양에 있어서 균사체 형태에 미치는 Ammonium Phosphate의 영향)

  • Lee, Kyu-Min;Lee, Shin-Young
    • The Korean Journal of Mycology
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    • v.29 no.2
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    • pp.91-98
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    • 2001
  • The mycelial morphology during submerged cultivation of Ganoderma ludium using by air-lift fermenter system were analyzed by image processing system and the characterization of mycelial morphology were investigated. In submerged culture using medium with different ammonium phosphate concentrations, the various morphological forms of G. lucidum mycelium were observed. The filamentous forms such as non-branched long filamentous mycelium, non-branched short mycelium, branched long filamentous mycelium, branched short mycelium, entangled mycelium and clump were observed, and also, and also, the pelleted forms such as smooth pellet, rough pellet and hollow rough pellet were observed. The mycelial morphology was changed from the filamentous to the pelleted forms by addition of ammonium phosphate. The fractal dimensions of pelleted and filamentous forms were 1.05 and 1.3, respectively, while the fractal dimension of mixtures of pelleted and filamentous forms was 1.16. Therefore, the fractal dimension was found to be more effective index for the detection of the mycelial morphology and morphological change during batch cultivation. The circularity was also found to be useful for evaluating the surface growth of pelleted mycelium.

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A study on the honeycomb entry and exit counting system for measuring the amount of movement of honeybees inside the beehive (벌통 내부 꿀벌 이동량 측정을 위한 벌집 입·출입 계수 시스템 연구)

  • Kim, Joon Ho;Seo, Hee;Han, Wook;Chung, Wonki
    • The Journal of the Convergence on Culture Technology
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    • v.7 no.4
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    • pp.857-862
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    • 2021
  • Recently, rapid climate change has had a significant impact on the bee ecosystem. The decrease in the number of bees and the change in the flowering period have a huge impact on the harvesting of beekeepers. Accordingly, attention is focused on smart beekeeping, which introduces IoT technology to beekeeping. According to the characteristics of beekeeping, it is impossible to continuously observe the beehive in the hive with the naked eye, and the condition of the hive is mostly dependent on knowledge from experience. Although a system that can measure partly through sensors such as temperature/humidity change inside the hive and measurement of the amount of CO2 is applied, there is no research on measuring the movement path and amount of movement of bees inside the beehive. Part of the migration of honeybees inside the hive can provide basic information to predict the most important cleavage time in beekeeping. In this study, we propose a device that detects the movement path of bees and measures and records data entering and exiting the hive in real time. The device proposed in this study was developed according to the honeycomb standard of the existing beehive so that beekeeping farms could use it. The development method used a photodetector that can detect the movement of bees to configure 16 movement paths and to detect the movement of bees in real time. If the measured honeybee movement status is utilized, the problem of directly observing the colony with the naked eye in order not to miss the swarming time can be solved.

The strengthening of democratic control over the authority of the superintendent of education in the Corona era

  • Park, Jong-Ryeol;Noe, Sang-Ouk
    • Journal of the Korea Society of Computer and Information
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    • v.26 no.6
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    • pp.145-154
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    • 2021
  • While welcoming the distributing of the powers of the Ministry of Education, which is currently being promoted, to the city and provincial offices of education, I am very concerned that this will expand and amplify the powers concentrated in one central institution to 17 local institutions closer to the field. Until now, the Ministry of Education and the Office of Education have served as co-cause providers as important reasons for hindering school education. The transfer of the authority of the Ministry of Education is highly likely to be deformed as soon as it results in the quantitative reduction of the functions and roles of the existing Ministry of Education and the quantitative expansion of the roles and functions of the city and provincial offices of education. In the reality that no legal device for school autonomy has been established, it is highly likely that emphasizing the principle of school autonomy in our educational climate, which is deeply rooted in the vertical bureaucratic administrative culture, will end with a simple measure or stop at the level of imitation. Therefore, a more stable device is needed to check the authority of the city and provincial office of education and the superintendent of education to take over the authority of the Ministry of Education. This is also a system that is still required even when school autonomy becomes legal. Therefore, it is necessary to revitalize the independent education committee, establish a local education committee in the city and province education office, and activate the resident participation system (resident proposal system, resident voting system, resident litigation system, resident audit request system).

In Vitro Nuclear Maturation of Canine Oocytes Obtained from Differents Stage of Estrus Cycle (개의 발정주기가 난자의 체외성숙에 미치는 영향)

  • Kim, Min-Kyu;Kim, Hye-Jin;Cho, Jong-Ki;Jang, Gu;Lee, Kyu-Seung;Kang, Sung-Geun;Lee, Byung-Chun;Hwang, Woo-Seok
    • Journal of Embryo Transfer
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    • v.17 no.2
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    • pp.145-151
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    • 2002
  • The aim of these experiments was to investigate in vitro nuclear maturation of canine oocyte collected from various stages of estrus cycles, Ovaries were obtained from 1 to 4 year-old mongrel bitch and minced for oocyte collection in phosphate buffered saline with 100 iu penicillin-G $m\ell$/sup -1/, 50 $\mu\textrm{g}$ streptomycin sulphate $m\ell$/sup -1/ and 0.1% (w/v) polyvinyl alcohol. Cumulus-oocyte-complexes (COCs) were washed in HEPES buffered tissue culture medium (TCM)199 and in vitro matured in TCM-199 culture medium supplemented with sodium pyruvate 0.028mg/$m\ell$, L-glutamine 0.146mg/$m\ell$, penicillin G 10,000 IU/$m\ell$, streptomycin 0.031mg/$m\ell$ and 10% (v/v) fatal calf serum. COCs were in vitro matured for 48~72 hrs at 39$^{\circ}C$ in humidified 5% $CO_2$ in air atmosphere. In vitro matured oocytes were remove the cumulus cells using 0.2% (v/v) hyaluronidase. After denuding, oocyte were placed in acetic acid : methanol : chlorform solusion (3:6 : 1.5 v/v) for 30 sec and acetic acid: ethanol(1:3 v/v) for 48hrs fixation. Nuclear maturation was classified to GV, GVBD, MI, MII and degenerate oocyte under microscopy after 1% aceto-orcein stain. In vitro maturation rates at 48hrs were not significantly difference among the oocytes collected from different stage of estrus at 15.9%, 16.3%, 23.7% and 18.2% for anestrus, proestrus, estrus and diestrus. However, the oocytes maturation(36.6%) of collected from estrus ovaries were significantly different from oocytes derived from proesturs, diestrus and anestrus ovaries(30.8%, 17.5% and 22.1%; p<0.05). The overall in vitro maturation rates were significantly higher (p<0.05) in 72hrs culture than 48hrs culture system. In summary, there was a tendency for higher in vitro maturation rates with the oocyte collected from estrus ovary than other stages of estrus. Also, for nuclear maturation, in vitro culture of oocyte for 72hrs was better than 48hrs culture.

Effects of Ectopic Expression of Transcription Factors on Adipogenic Transdifferentiation in Bovine Myoblasts (한우(Bos taurus coreanae) 유래 myoblast에서 전사인자 과발현에 의한 지방세포로의 교차 분화 유도)

  • Moon, Yang Soo
    • Journal of Life Science
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    • v.22 no.10
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    • pp.1316-1323
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    • 2012
  • The present study was conducted to investigate whether myoblasts can be transdifferentiated into adipocytes by ectopic expression of adipogenic transcription factors, including peroxisome proliferator-activated receptor-${\gamma}$ ($PPAR{\gamma}$), CCAAT/enhancer-binding protein-${\alpha}$ (C/$EBP{\alpha}$), sterol regulatory element binding protein-1c (SREBP1c), and Krueppel-like factor 5 (KLF5), in primary bovine satellite cells. Transcription factors were transiently transfected into primary bovine myoblasts, and the cells were cultured with adipogenic differentiation medium for 2 days and then cultured on growth medium for an additional 8 days. Ectopic expression of $PPAR{\gamma}$ or C/$EBP{\alpha}$ alone was insufficient to induce adipogenesis in myoblasts. However, overexpression of both $PPAR{\gamma}$ and C/$EBP{\alpha}$ in myoblasts was able to induce adipogenic transdifferentiation as indicated by the appearance of mature adipocytes, the induction of adipogenic gene expressions, and the suppression of myogenic gene expressions. In addition, KLF5 and $PPAR{\gamma}$ co-transfected bovine myoblasts were converted to adipocytes but not in cells transfected with only KLF5 expression vector. Overexpression of SREBP1c alone was sufficient to induce transdifferentiation from myoblasts into adipocytes. These results demonstrate that primary bovine satellite cells can be transdifferentiated into adipocytes either by single ectopic expression or combined expression of adipogenic transcription factors in a culture system.

Characteristics of ${\beta}$-Glucosidase Secreted by Trichoderma reesei KCTC 6952 (Trichoderma reesei KCTC 6952로부터 분비된 ${\beta}$-glucosidase의 특성)

  • Park, Sung-Hee;Oh, Min-Jung;Lee, Jeong-Rai;Kwon, Suk-Hyung;Choi, Young-Wook;Lee, Min-Won;Kim, Keun-Sung
    • Korean Journal of Food Science and Technology
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    • v.37 no.3
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    • pp.504-507
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    • 2005
  • Trichoderma reesei KCTC 6952 possesses cellulase system consisting of three enzymatic components necessary to synergistically hydrolyze crystalline cellulose, among which ${\beta}$-glucosidase effectively releases glucose from glycoside derivatives. ${\beta}$-Glucosidase of T. reesei KCTC 6952 grown in modified Mandels' medium showed maximum activity(1.33 unit/mL) 4 days after initiation of growth. Optimal reaction conditions of the enzyme were 50 mM sodium acetate (pH 5) at $70^{\circ}C$ for 10 min. Enzymatic activities stabilized below $50^{\circ}C$ at pH range of 4-5. Results show ${\beta}$-glucosidase exerted its catalytic activities at relatively high temperatures and broad pH range.

Exploration of suitable rice cultivars for close mixed-planting with upland-adapted cereal crop

  • Shinohara, Nodoka;Shimamoto, Hitoshi;Kawato, Yoshimasa;Wanga, Maliata A.;Hirooka, Yoshihiro;Yamane, Koji;Iijima, Morio
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.304-304
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    • 2017
  • In semi-arid countries such as Namibia, the flooding unexpectedly happens in a rainy season, causing losses in the yield of upland-adapted cereal crop. In flooding conditions, rice roots sequentially form aerenchyma and a barrier to radial oxygen loss (ROL), and oxygen is released into the rhizosphere near the root tips. Iijima et al. (2016) and Awala et al. (2016) reported that close mixed-planting with rice can mitigate the flood stress of co-growing upland-adapted cereal crop by modifying their rhizosphere microenvironments via the oxygen released from the rice roots. Moreover, by using the model system of hydroponic culture, it was confirmed that oxygen from rice roots was transferred to co-growing upland-adapted cereal crop in close mixed planting system (Kawato et al., 2016). However, it is not sure whether the ability of oxygen release varies among rice cultivars, because Kawato et al. (2016) used only one japonica cultivar, Nipponbare (Oryza sativa). The objective of this study was to compare the ability of oxygen release in rhizosphere among rice cultivars. The experiment was conducted in a climate chamber in Kindai University. We used 10 rice cultivars from three different rice species (O. sativa (var. japonica (2), var. indica (3)), Oryza glaberrima Steud. (2) and their interspecific progenies (3)) to compare the ability of oxygen release from the roots. According to the method by Kawato et al. (2016), the dissolved oxygen concentration of phase I (with shoot) and phase II (without shoot) were measured by a fiber optic oxygen-sensing probe. The oxygen released from rice roots was calculated from the difference of the measurements between phase I and phase II. The result in this study indicated that all of the rice cultivars released oxygen from their roots, and the amount of released oxygen was significantly correlated with the above-ground biomass (r = 0.710). The ability of oxygen release (the amount of the oxygen release per fresh root weight) of indica cultivars (O. sativa) tended to be higher as compared with the other cultivars. On the other hand, that of African rice (O. glaberrima) and the interspecific progenies tended to be lower. These results suggested that the ability of oxygen release widely varies among rice cultivars, and some of indica cultivars (O. sativa) may be suitable for close mixed-planting to mitigate flood stress of upland-adapted cereal crop.

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Optimization of In Vitro Culture System of Mouse Preantral Follicles (생쥐 Preantral Follicles의 체외 배양 시스템에 관한 연구)

  • Park, Eun-Mi;Kim, Eun-Young;Nam, Hwa-Kyung;Lee, Keum-Sil;Park, Sae-Young;Yoon, Ji-Yeon;Heo, Young-Tae;Cho, Hyun-Jung;Park, Se-Pill;Loo, Jin-Ho
    • Clinical and Experimental Reproductive Medicine
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    • v.28 no.2
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    • pp.95-103
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    • 2001
  • 연구목적: 본 연구는 생쥐 preantral follicles의 체외 배양 조건을 확립하고 이를 기초로 높은 체외 발달률 그리고 산자 생산률을 얻고자 하였다. 연구재료 및 방법 : Preantral follicles의 oocyte-granulosa cell complexes (OGCs)는 생후 12일된 FI ($C57BL{\times}CBA$)으로부터 난소를 적출하여 효소를 이용한 방법을 통해 획득하였다. 회수된 complexes는 10일 또는 12일 동안 체외 성장을 위해 Transwell-COL membrane insert로 옮겨졌고 5% FBS, 100 mIU/ml FSH, 100 mIU/ml hMC가 첨가된 ${\alpha}MEM$에서 배양되었다. 체외 성숙을 위해 1.5 IU/ml hCG가 첨가된 ${\alpha}MEM$에서 18 hrs 배양을 실시하였다. 그 후 M16에서 수정능력이 획득된 정자와 수정을 하여 4 hrs, 7 hts, 9 hrs 후에 10% FBS가 첨가된 modified M16 배양액에서 4일간 배양하거나 또는 bovine cumulus cell과 co-culture를 실시하였다. 그리고 형태적으로 정상적인 22개의 상실배와 포배를 2마리의 위임신 대리모 (ICR)의 자궁에 이식하여 산자 생산을 유도하였다. 결과: 1) OGCs 크기가 mouse preantral follicles의 핵 및 세포질 성숙에 미치는 영향을 조사하였을 때 $120{\sim}150{\mu}m$의 preantral follicles (MII: 33.0%, 난할률: 36.7%, 상실배 이상; 20.9%)은 핵 및 세포질 성숙에 있어서 $70{\sim}110{\mu}m$ (MII: 12.2%, 난할률: 10.2%, 상실배 이상: 4.8%)보다 더 높았다(p<0.001). 2)체외 성장기간의 연장이 mouse preantral follicles의 핵 및 세포질 성숙에 미치는 영향을 조사하였을 때 10일 (난할률: 38.2%)은 12일 (난할률: 20.0%)보다 난할률에서만 더 높았다 (p<0.01). 3) 체외 수정 시간의 연장이 mouse preantral follicle의 세포질 성숙에 미치는 영향을 조사하였을 때 9 hrs (난할룰 31.5%, 상실배 이상: 14.3%)은 4 hrs (난할률: 17.5%, 상실배 이상: 4.8%), 7 hrs (난할률: 20.4%, 상실배 이상: 6.1%) 보다 세포질성숙에 있어서 유의하게 높은 발달률을 나타냈다 (p<0.01). 4) 공배양이 mouse preantral follicle의 세포질성숙에 미치는 영향을 조사하였을 때 공배양 (상실배 이상: 17.4%)을 실시했을 때와 M16 (상실배 이상17.4%)에서 배양되었을 때는 차이가 없었다. 5)preantral follicle의 크기 ($120{\sim}150{\mu}m$), 체외 성장기간 (10일), 체외 수정 기간 (9시간), 배양 환경 (단지 medium만 존재)의 적절한 결과들을 종합하여 수행하였을 때 MII 성숙률, 난할률, 상실배 이상의 발달률은 30.2%, 39.3%, 22.5%이었고 총 22개의 상실배 및 포배를 2마리의 대리모에 이식했을 때 1마리가 임신했고 1마리의 산자를 생산했다. 결론: 따라서, 본 실험은 preantral follicle을 이용한 체외 배양 시스템이 생쥐 oocyte를 공급하는 또 다른 방법으로 효과적으로 이용될 수 있다는 것을 시사한다.

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