• Title/Summary/Keyword: 세포 계수기

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Derivation of Mouse ES Cells from Isolated Blastomeres in Culture Media Supplemented with LIF (LIF를 첨가한 배양액을 이용한 할구 유래 생쥐 배아줄기세포주의 확립)

  • Cho, Jae-Won;Lim, Chun-Kyu;Ko, Duck-Sung;Kang, Hee-Jung;Jun, Jin-Hyun
    • Development and Reproduction
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    • v.12 no.1
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    • pp.77-86
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    • 2008
  • This study was carried out to investigate the effect of leukemia inhibitory factor (LIF) on the derivation of mouse ES cells from isolated blastomeres. Two-cell stage mouse embryos were obtained from superovulated BDF1 female mice. Collected embryos were cultured to blastocyst stage in culture medium supplemented with 0, 1,000, 2,500 or 5,000 U/mL of LIF. Cultured blastocysts were examined by counting the number of cells in the inner cell mass (ICM) and trophectoderm (TE) using differential staining method. When 2-cell embryos were cultured with 2,500 U/ml of LIF, the cell numbers of ICM significantly increased in comparing with those of the control($21.0{\pm}4.0$ vs. $15.9{\pm}5.0$, P<0.01) and 1,000 U/mL of LIF-containing group ($21.0{\pm}4.0$ vs. $16.6{\pm}4.9$, P<0.05). We used an ES cell establishment medium with 20% Knockout Serum Replacement and 0.01 mg/mL ACTH instead of fetal bovine serum. Establishing efficacy of ES cell lines were the highest in 2,500 U/mL of LIF-containing group as 36.7% (11/30). This culture medium was applied to the culture of isolated blastomeres and to derivate ES cell lines. Three ES cell lines (21.4%) from isolated blastomeres of 2-cell stage embryos were established. In further experiments, we could establish one ES cell line (4.0%) from single blastomere of 4-cell stage embryo. The subcultured ES cells and their embryoid bodies were characterized by analyzing gene expression for undifferentiation and differentiation marker gene using immunocytochemistry and RT-PCR. In conclusion, LIF supplementation in culture medium could increase the cell number in ICM of blastocysts and support derivation of ES cell lines from isolated blastomeres.

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Utilization of Obosan (Dietary Herbs) I. Effects on Survival, Growth, Feed Conversion Ratio and Condition Factor in Olive Flounder, Paralichthys olivaceus (한방사료 첨가제인 어보산의 효과 I. 넙치의 생존율, 성장, 사료효율 및 비만도에 미치는 영향)

  • 김동수;김종현;정창화;이상윤;이상민;문영봉
    • Journal of Aquaculture
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    • v.11 no.2
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    • pp.213-221
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    • 1998
  • The effects of differen concentrations of Obosan as a feed additive dietary herb were examined on survival rate, growth, feed conversion ration and condition factor in olive flounder (paralichthys olivaceus). Effectiveness of dietary Obosan with optimized concentration for 48 weeks were also observed with regard to growth performances and yields. All groups fed diets containing 0.15, 0.3 and 0.6% of Obosan revealed significantly higher survival rate than control group (P<0.05). Growth, feed conversion ratio and condition factor of olive flounder fed diets containing Obosan were considerably improved when compared to those of controls (P<0.05). The 0.3% of dietary Obosan was proven to be the optimal concentration in all parameters tested. The dietary Obosan (0.3%) for 48 weeks showed significantly higher surval rate than control (P<0.05), and also improved yields in weight gain (19.0% improvement), specific growth rate (4.8%), feed conversion ratio(13.6%) and condition factor (10.8%), significantly (P<0.05).

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Effect on the Production of Toxin Protein of Bacillus thuringiesis by Culture Conditions (배양 조건이 Bacillus thuringiesis 독소단백질의 생산에 미치는 영향)

  • Kim, Tae Wan;Kim, Tae Hwan;Yu, Yong Man
    • The Korean Journal of Pesticide Science
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    • v.18 no.4
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    • pp.358-363
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    • 2014
  • We was investigated parasporal inclusion proteins change to use industrial medium of new strain Bacillus thuringiensis CAB 565, CAB 566. To confirm medium's oxygen efficient consist of glucose and yeast extract, we was conducted oxygen transfer coefficients (KLa) of medium's concentration and impeller in 20 l-Jar fermentor. When to increase air flow rate and medium concentration, KLa rate is rise. Also it is effective on agitation rate 200 rpm, but KLa rate is decrease when to rise agitation rate. To hold dissolved oxygen rate (upper 50%), Air flow rate is steadily increase on culture to use microsparger. When 16 hour of culture stage, B.t. CAB 565 and B.t. CAB 566 harvested respectively $2.3{\times}10^{10}$, $1.8{\times}10^{10}$ viable cell/ml. When 54 hour, B.t. CAB565, 566 harvested respectively $1.9{\times}10^{10}$, $1.4{\times}10^{10}spore/ml$. To resulting carbon's concentration, It is the most effective that glucose concentration is contained 5% in medium.

Studies on Whole Cell Immobilized Glucose Isomerase - II. Operational Studies on the Batchwise and Continuous Isomerization of D-Glucose - (포도당 이성화 효소의 세포 고정화에 관한 연구 - 제 2 보 : 회분식 및 연속 반응조를 사용한 포도당의 이성화 -)

  • Ahn, Byung-Yoon;Byun, Si-Myung
    • Korean Journal of Food Science and Technology
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    • v.11 no.4
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    • pp.249-257
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    • 1979
  • Using the whole cell immobilized glucose isomerase which was prepared in the previous work (Korean J. Food Sci. & Technol., 11(3), 192 (1979), the specific activity of the immobilized enzyme was 48.1 units in the batch reaction system and 114 units in the continuous reaction system per g of matrix, respectively. In the continuous reactor the voidity was 0.36, which was suitable for the packed bed reactor. This immobilized enzyme showed a good operational stability of 115 days of half life which was sufficient for the continuous operation. The experimental result showed that 55 % of the substrate was converted to the product in the packed bed reactor. The productivity was dependent on the flow rate, column geometry, enzyme loading, and substrate concentration. An intrapaticle diffusion was observed by the effectiveness factor of 0.75 and interparticle diffusion by the decrease of Km' with increasing the superficial velocity.

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Fabrication of a Micro-thermoelectric Probe (마이크로 프로브 기반 열전 센서 제작 기술)

  • Chang, Won-Seok;Choi, Tae-Youl
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.35 no.11
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    • pp.1133-1137
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    • 2011
  • A novel technique for the fabrication of a glass micropipette-based thermal sensor was developed utilizing inexpensive thermocouple materials. Thermal fluctuation with a resolution of ${\pm}0.002$ K was measured using the fabricated thermal probe. The sensors comprise unleaded low-melting point solder alloy (Sn) as a core metal inside a borosilicate glass pipette coated with a thin film of Ni, creating a thermocouple junction at the tip. The sensor was calibrated using a thermally insulated calibration chamber, the temperature of which can be controlled with a precision of ${\pm}0.1$ K and the thermoelectric power (Seebeck coefficient) of the sensor was recorded from 8.46 to $8.86{\mu}V$/K. The sensor we have produced is both cost-effective and reliable for thermal conductivity measurements of micro-electromechanical systems (MEMS) and biological temperature sensing at the micron level.

Surface Modification by Laser Deposition and Femtosecond Laser for Biomedical Applications (레이저증착과 펨토레이저를 이용한 생체의료분야의 표면처리응용)

  • Choe, Han-Cheol
    • Proceedings of the Korean Institute of Surface Engineering Conference
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    • 2015.05a
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    • pp.24-24
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    • 2015
  • 최근 생체재료의 개발이 눈부시게 발전되고 생체적합성이 우수한 표면을 요구함에 따라 생체재료의 표면처리에 대한 연구가 활발히 진행되고 있다. Laser Deposition법은 항공기 부품제조 분야에 주로 사용되고 있으며 최근에 오하이오 주립대 타이타늄합금연구센터를 중심으로 표면처리에 관한 연구가 주로 이루어졌다. 특히 이를 이용하여 치과재료의 표면처리에 응용을 시도하였다. 치과에서 응용될 수 있는 경우는 주로 임플란트는 부분 또는 완전 무치악 환자의 보철수복에 사용되는 보철물의 제작등에 사용될 수 있으며 이중에서도 특히 생체용 임플란트의 표면처리응용으로 임플란트와 조직간의 접합성을 개선하는 표면처리법으로 연구되었다. 임플란트의 성공과 실패는 물성적인 측면에서 임플란트의 형태, 표면거칠기 및 표면처리방법, 초기하중 등에 의하여 좌우되며 임플란트 재료에 작용하는 응력차폐는 생체적합성을 좌우하는 큰 요인이 되고 있다. 이를 위하여 저 탄성계수합금을 설계하지만 하중을 버티는 강도가 낮아지는 단점이 있어 레이저증착법을 이용하여 임플란트재료인 Ti6Al4V합금에 탄성계수가 낮은 Ta, Nb등을 코팅하는 방법을 통하여 이를 해결하고자하는 시도가 이루어지고 있다. 이 방법은 최근의 3D 프린팅의 원리가 되고 있다. 따라서 발표에서는 Laser Deposition방법을 이용하여 치의학분야에서 응용되고 있는 예를 강연하고 응용 가능 분야에 대하여 토론 하고자한다. 또한 펨토레이저를 이용하여 생체합금의 표면처리는 생체활성화를 더욱 증진시키며 이를 위하여 많은 연구 수행되고 있다. 본 발표에서는 매식용 합금 표면에 펨토레이저를 이용하여 텍스춰링하여 세포가 잘 성장 할 수 있는 크기의 조절함으로써 기존의 표면처리와는 다른 효과를 얻을 수 있는 장점을 알아본다. 펨토레이저를 이용하면 여러 가지 형태의 텍스춰링이 가능하며 원형, 사각형등등 자유자제로 형태의 묘사가 가능하고 깊이 또한 쉽게 조절할 수 있는 장점이 있다. 지금까지는 표면 개질에 사용되는 레이저는 주로 Nd:YAG 레이저의 파장을 반으로 줄인 녹색레이저 (${\lambda}=532nm$)를 사용하거나, 자외선파장영역의 레이저를 사용하는 경우가 일반적으로 가장 보편화되었다. 이를 이용하여 제조된 Ti합금에 펨토 초(10-15 second) 펄스폭 대역을 갖는 레이저를 이용하여 나노크기의 미세 요철을 표면에 형성한 후, 나노튜브를 형성하여 그 표면특성의 변화를 알아보고 펨토레이저가 의료분야에 적용되고 있는 예를 살펴보고자 한다.

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Studies on Genetics and Breeding of Rainbow Trout(Oncorhynchus mykiss) II. Relationships between Levels of Serum LH, FSH and Estradiol-$17{\beta}$ ; and Levels of Albumin, Total Protein, and Triglycerides ; and their Effect on the Changes of Gonadosomatic Indices in the Ovarian Development in the Rainbow Trout (무지개송어의 유전 육종학적 연구 II. 무지개송어의 난소 발달에 영향을 미치는 혈청중 LH, FSH, Estradiol-$17{\beta}$ 수준과 Albumin, Total Protein, Triglycerides 함량과의 관계 및 성선지수 변화간의 상관관계)

  • Yoon Jong-Man;KIM Gye-Yoong;PARK Hong-Yang
    • Journal of Aquaculture
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    • v.4 no.2
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    • pp.97-110
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    • 1991
  • This study was undertaken to understand the seasonal changes of hormone levels such as luteinizing hormone (LH), follicle stimulating hormone (FSH) and estradiol-$17{\beta}$, serum components such as albumin, total protein and triglycerides, and GSI of rainbow trout (Onco-rhynchus mykiss) under the normal-light circumstances. LH levels were the highest in November whereas FSH levels were decreased progressively from October to February. Serum GTH, such as LH and FSH, levels were low immediately after egg-stripping and were significantly increased in the summer in association with the initiation of vitellogenesis. Serum albumin, total protein, and triglycerides were significantly increased in August in association with initiation of vitellogenesis and were peak in November prior to egg-stripping. Oocyte growth, measured by oocyte diameter and GSI, was significantly increased in August and showed the highest value in January. Correlation coefficients of serum LH with FSH and estradiol-$17{\beta}$ were +0.844 and +0.947, respectively. Correlation coefficients of estradiol-$17{\beta}$ with albumin and total protein were +0.634 and +0.859, respectively. Correlation coefficients of serum estradiol-$17{\beta}$ with triglycerides and GSI were +0.673 and +0.694, respectively. Probably by positive feedback mechanism, LH, FSH and estradiol-$17{\beta}$ appeared to stimulate to liver to secrete albumin, total protein and triglycerides for vitellosenesis.

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Reproductive Ecology of the Freshwater Marsh Clam, Corbicula papyracea (Heude) in Bukhan River (북한강 엷은재첩, Corbicula papyracea (Heude) 의 번식생태)

  • Kim, Dae-Hee;Lee, Wan-Ok;Lee, Jun-Sang
    • The Korean Journal of Malacology
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    • v.30 no.2
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    • pp.107-116
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    • 2014
  • Gonadal development and maturation mechanism were studied on the freshwater marsh clam Corbicula papyracea (Heude), which is the endangered species in Korea. The specimens were collected in the rearing ponds and waterway of NFRDI in Gapyeong-gun, Gyeonggi-do from January to December 2004, and then investigated by condition factor, relative growth, gonadal development phases and gonad histological characters based on 30-50 individuals every month. Comparing with the freshwater marsh clam, C. papyracea is small, light olive brown shell and violet interior. The hermaphrodite individuals of C. papyracea take an internal fertilization and fertilized eggs are stored in the foster-sack in the gills, then the hatched juveniles are released outside after an ovoviviparous process. The average water temperature of inhabit area was in range of $1.8-27.0^{\circ}C$ and usually took great effects on the gonad maturation of C. papyracea. The condition factor ranged from 0.14 to 0.21 throughout the year, which was the lowest during winter season (December-February), and gradually increased to the highest value of 0.21 in May. The ratio of meat weight to total weight was 25.9-38.7%, indicating the similar trend with condition factor. The highest values of condition factor and the ratio of meat weight appeared 1-2 months later than gonadosomatic index reached the peak value, it was probably because that the ovoviviparous eggs would spend a long period before hatching from the foster-sack in the gills. To synthesize the characters of meat weight, condition factor and gonad development by histological study, reproductive cycle of C. papyracea could be divided into five successive stages: multiplicative stage (December to February), growing stage (February to May), mature stage (June to August), spawning stage (August to November), recovery stage (November to December). The smallest shell length of matured C. papyracea was 12.6 mm, and individuals, larger than 16 mm, was formed the nursery in the gills.

Interspecific Variation in the Protoplast Formation of the Genus Cellulomonas (Cellulomonas속 종간의 원형질체 형성조건의 차이에 대하여)

  • Lee, Eun-Ju;Bae, Moo
    • Korean Journal of Microbiology
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    • v.24 no.2
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    • pp.154-160
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    • 1986
  • In order to develope interspecific fusion of the genus Cellulomonas capable of assimilation cellulose, the optimun conditions for the protoplast formation was investigated to examine the susceptibility of cell wall, between different species of the same genus using scanning electron microscope. The variation in the susceptibilities of Cellulomonas sp. CS 1-1 and C. flavigena to lysozyme treatment were considerably remarkable, although they belong to the same genus. The rate of protoplast formation of CS1-1 was 99.9% being treated with lysozyme $(100{\mu}g/ml)$ for 30 minute and that of C. flavigena was about 80% being treated at the concentration of $600{\mu}g/ml$ of lysozyme for 6 hours. The susceptibility of cell wall to the lysozyme treatment on protoplast formation of the strain, CS1-1 seems not to be depend on the cultural periods of cells. On the contrary, that of C. flavigena was considerably depend on the periods. Cells of C. flavigena at mid exponential phase could be more efficiently converted to protoplast cells than those at late exponential phase be done. The rate of the protoplast formation was 95%, when cells of C. flavigena at mid exponential phase were treated with lysozyme $600{\mu}g/ml$ for 6 hours and observed by SEM. In the evalution of protoplast formation of the CS1-1 results of counting method in plate after osmotic shock treatment were similar to the results of the direct observation method by means of SEM. But in the case of C. flavigena the latter method was much more reliable than the former, because the differences between the number of spheroplasts and protoplasts were not able to figure out on conuting the number of protoplast after osmotic shock tretment.

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The Study of CYFRA 21-1 and Epidermal Growth Factor Receptor Levels in Cancer Tissue of Bronchogenic Carcinoma Patients (폐암환자의 암조직내 CYFRA 21-1과 Epidermal Growth Factor Receptor의 측정치에 대한 연구)

  • 김대연;김송명
    • Journal of Chest Surgery
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    • v.30 no.9
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    • pp.854-861
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    • 1997
  • CYPRA 21-1 is known to be a cytokeratin 19 fragment, and it can be detected by using two specific monoclonal antibodies (KS 19-1 and BM 19-21) and can be clinically applied as a useful circulating tumor marker The epidermal growth factor receptor (EGF-R) expression was evaluated and characterized by its tyrosine protein kinase activity and by its ligand-stimulated autophosphorylation, a property shared with other peptide growth factor receptors. Autocrine or para'urine action was initiated by a growth factor, or by a transforming growth factor o, which had an extensive homology with EGP and which also stimulated tyrosine kinase activity on the EGF-R. The CYFRA 21-1 and the EGF-R levels in 30 patients with primary lung tumors were investigated. There were 24 patients with squamous cell carcinomas and 6 patients with adenocarcinomas. Specimen 5 mm3 in size were sampled at three different locations ; the main lesion, the boundary between the lesion and the unaffected tissue, and the unaffected tissue of the patients. The results were as follows 1. The CYPRA 21-1 concentration in the cancer boundary, the most malignant region,(348.6 : 89.9 ng/ml) was the lowest value. The CYFRA 21-1 concentration in unaffected tissue,(718.4$\pm$77.8 ng/ml) was higher than that in the main lesion. which had intact cellularity. 2. The EGF-R concentration in the main lesion was higher than that in the unaffected tissue, and the EGF-R concentration in a squamous cell cacinoma was higher than that in an adenocarcinoma. also, the EGF-R concentration in the cancer b undary was highest at stage 1, ll. The EGF-R concentration was higher in the main cancer lesion that in the unaffected tissue at stage 111, IV. 3. The CYFRA 21-1 was a cytoplasmic skeleton and the EGF-R was a cell-wall component; there was no correlation. In conclusion, CYFRA 21-1 was abundant in the cytoplasm but had a higher concentration in the unaffected tissue than in the main cancer lesion. The CYFRA 21-1 concentration of the tissue did not reflect the amount of cancer activity, the EGP-R was located in the cell membrane, the level of tissue that reflects cancer activity, so the main cancer lesion had a higher concentration than the unaffected tissue. CYFRA 21-1 is not a useful tumor maker at the tissue level. Because the EGF-R concentration re(looted the cancer activity, its a useful tumor marker for lung cancer.

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