• 제목/요약/키워드: In-vitro techniques

검색결과 351건 처리시간 0.031초

Effect of MgO-P2O5 Sintering Additive on Microstructure of Sintered Hydroxyapatite (HAp) Bodies and Their In-Vitro Study

  • Lee, Byong-Taek;Youn, Hyeong-Chul;Lee, Chi-Woo;Song, Ho-Yeon
    • 한국재료학회지
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    • 제17권2호
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    • pp.100-106
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    • 2007
  • The effects of $MgO-P_2O_5$ based sintering additive on the microstructure and material and biological properties of hydroxyapatite $(HAp,\;Ca_{10}(PO_4)_6(OH)_2)$ ceramic were investigated using XRD, SEM and TEM techniques. The $MgO-P_2O_5$ sintering additive improved the material properties and increased the grain size in the sintered HAp bodies. As the content of sintering additive increased over 4 wt%, a small amount of the HAp phase was decomposed and transformed to ${\beta}-TCP$. In the 2 wt% $MgO-P_2O_5$ content HAp sintered body, the maximum values of density and hardness were respectively about 3.10 gm/cc and 657 HV. However, the maximum fracture toughness in the HAp body containing 8 wt% $MgO-P_{2}O_{5}$ was about $1.02MPa{\cdot}m^{1/2}$ due to the crack deflection effect. Human osteoblast like MG-63 cells and osteoclast like raw 264.7 cells were well grown and fully covered all of the HAp sintered bodies. The osteoblast cells were grown with spindle-shaped and the osteoclast cells had a grape-like round shape.

Efficient Derivation of New Human Embryonic Stem Cell Lines

  • Kim, Sun Jong;Lee, Jeoung Eun;Park, Jong Hyuk;Lee, Jung Bok;Kim, Jin Mee;Yoon, Byung Sun;Song, Ji Min;Roh, Sung Il;Kim, Chul Geun;Yoon, Hyun Soo
    • Molecules and Cells
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    • 제19권1호
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    • pp.46-53
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    • 2005
  • Human embryonic stem (hES) cells, unlike most cells derived from adult or fetal human tissues, represent a potentially unlimited source of various cell types for basic clinical research. To meet the increased demand for characterized hES cell lines, we established and characterized nine new lines obtained from frozen-thawed pronucleus-stage embryos. In addition, we improved the derivation efficiency from inner cell masses (to 47.4%) and optimized culture conditions for undifferentiated hES cells. After these cell lines had been maintained for over a year in vitro, they were characterized comprehensively for expression of markers of undifferentiated hES cells, karyotype, and in vitro/in vivo differentiation capacity. All of the cell lines were pluripotent, and one cell line was trisomic for chromosome 3. Improved culture techniques for hES cells should make them a good source for diverse applications in regenerative medicine, but further investigation is needed of their basic biology.

Growth of Endothelial Cells on Microfabricated Silicon Nitride Membranes for an In Vitro Model of the Blood-brain Barrier

  • Harris, Sarina G.;Shuler, Michael L.
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제8권4호
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    • pp.246-251
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    • 2003
  • The blood-brain barrier (BBB) is composed of the brain capillaries, which are lined by endothelial cells displaying extremely tight intercellular junctions. Several attempts at creating an in vitro model of the BBB have been met with moderate success as brain capillary endothelial cells lose their barrier properties when isolated in cell culture. This may be due to a lack of recreation of the in vivo endothelial cellular environment in these models, including nearly constant contact with astrocyte foot processes. This work is motivated by the hypothesis that growing endothelial cells on one side of an ultra-thin, highly porous membrane and differentiating astrocyte or astrogliomal cells on the opposite side will lead to a higher degree of interaction between the two cell types and therefore to an improved model. Here we describe our initial efforts towards testing this hypothesis including a procedure for membrane fabrication and methods for culturing endothelial cells on these membranes. We have fabricated a 1 $\mu\textrm{m}$ thick, 2.0 $\mu\textrm{m}$ pore size, and 55% porous membrane with a very narrow pore size distribution from low-stress silicon nitride (SiN) utilizing techniques from the microelectronics industry. We have developed a base, acid, autoclave routine that prepares the membranes for cell culture both by cleaning residual fabrication chemicals from the surface and by increasing the hydrophilicity of the membranes (confirmed by contact angle measurements). Gelatin, fibronectin, and a 50/50 mixture of the two proteins were evaluated as potential basement membrane protein treatments prior to membrane cell seeding. All three treatments support adequate attachment and growth on the membranes compared to the control.

호르몬 및 배지 고형제를 이용한 알스트로메리아 식물체 대량증식 체계 확립 (Establishment of propagation system for Alstroemeria plants by using hormones and gelling agents)

  • 양환래;이상희;김종보
    • 문화기술의 융합
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    • 제4권4호
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    • pp.167-171
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    • 2018
  • 알스트로메리아는 경제적으로 중요한 절화 작물 중 하나이다. 그러나 낮은 증식률과 번식기간이 길고 바이러스 질병 감염이 높기 때문에 대량증식이 어렵다고 알려져 있다. 따라서 기내 조직배양을 이용한 대량증식 시스템 쳬계 확립이 필요하다. 본 연구에서는 BA와 NAA 그리고 여러 가지 배지 고형제를 이용하여 알스트로메리아 최적의 생장 효율을 찾기 위하여 실험을 진행하였다. 먼저 BA 1.0 mg/L 와 NAA 0.1 mg/L를 첨가한 배지에서 무처리구에 비하여 신초 및 뿌리의 개수가 약 1.5배 증가하였고 비대한 신초와 뿌리를 나타내었다. 여러 가지 배지 고형제들 중 gelrite 0.25%를 첨가한 배지에서 타 처리구에 비해 최대 50% 향상된 길이를 나타내었고, 근경 개수 및 생체중 증가량에 있어서도 가장 높은 효율을 나타내었다. 본 연구결과를 토대로 알스트로메리아 신품종 개발 및 대량증식 시스템 체계 확립에 도움을 줄 것으로 기대된다.

생쥐 정자세포의 분리와 체외 배양에 관한 연구 (In Vitro Culture Following Purfication of Mouse Spermatogenic Cells)

  • 김묘경;김진회;이상민;정/하해숙;이훈택;정길생
    • 한국가축번식학회지
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    • 제20권1호
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    • pp.43-52
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    • 1996
  • 본 연구는 생쥐의 고게정관내에 존재하는 분화단계의 정자세포를 발생단계별로 분리하여, 체외에서 단기간 배양체계를 확립하기 위하여 실시하였다. 8주령 이상된 생쥐의 정소로부터 정소막을 제거시킨 후, collagenase (1mg/ml), trypsin(2.5mg/ml)를 처리하여 곡세정관을 간질세포와 분리하여 배양액에 부유시켰다. 부유세포는 Celcep장치를 이용하여 세포크기와 밀도 차이에 의해 분화 단계별로 분리하였다. 회수된 세포의 균질성은 haematoxylin/eosin 또는 orcein으로 염색한 후, 광학현미경하에서 확인한 결과 약 85% pachytene spermatocyte와 round spermatid을 성공적으로 분리해냈다. 따라서 sedimentation velocity에 의해서 생쥐의 spermatogenic cell의 발달단계별 분리가 가능함을 알 수 있었다. 이러한 방법으로 분리된 pachytene spermatogenic cell들은 DMEM 배양액에서 6일 이상 배양한 결과 약 36%의 생존율을 보였다. 따라서, 분화단계별 정자 세포의 분리 및 배양체계의 확립은 웅성생식세포의 발생과정에 따른 생리 또는 분자생물학적 현상을 규명함은 물론 세포융합기술의 이용에 의한 형질전환동물 생산에의 응용을 통해 가축에 있어서의 형질전환 생산효율의 개선에 기여할 수 있으리라 사료된다.

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PIV System for the Flow Pattern Anaysis of Artificial Organs ; Applied to the In Vitro Test of Artificial Heart Valves

  • Lee, Dong-Hyeok;Seh, Soo-Won;An, Hyuk;Min, Byoung-Goo
    • 대한의용생체공학회:의공학회지
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    • 제15권4호
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    • pp.489-497
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    • 1994
  • The most serious problems related to the cardiovascular prothesis are thrombosis and hemolysis. It is known that the flow pattern of cardiovascular prostheses is highly correlated with thrombosis and hemolysis. Laser Doppler Anemometry (LDA) is a usual method to get flow pattern, which is difficult to operate and has narrow measure region. Particle Image Velocimetry (PIV) can solve these problems. Because the flow speed of valve is too high to catch particles by CCD camera, high-speed camera (Hyspeed : Holland-Photonics) was used. The estimated maximum flow speed was 5m/sec and maximum trackable length is 0.5 cm, so the shutter speed was determined as 1000 frames per sec. Several image processing techniques (blurring, segmentation, morphology, etc) were used for the preprocessing. Particle tracking algorithm and 2-D interpolation technique which were necessary in making gridrized velocity pronto, were applied to this PIV program. By using Single-Pulse Multi-Frame particle tracking algorithm, some problems of PIV can be solved. To eliminate particles which penetrate the sheeted plane and to determine the direction of particle paths are these solving methods. 1-D relaxation fomula is modified to interpolate 2-D field. Parachute artificial heart valve which was developed by Seoul National University and Bjork-Shiely valve was testified. For each valve, different flow pattern, velocity profile, wall shear stress and mean velocity were obtained.

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The effect of embryo catheter loading technique on the live birth rate

  • Omidi, Marjan;Halvaei, Iman;Mangoli, Esmat;Khalili, Mohammad Ali;Razi, Mohammad Hossein
    • Clinical and Experimental Reproductive Medicine
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    • 제42권4호
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    • pp.175-180
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    • 2015
  • Objective: Embryo loading (EL) is a major step in embryo transfer (ET) and affect on the success of in vitro fertilization (IVF). This study aimed to compare the effect of two different EL techniques on the rates of pregnancy and delivery in IVF/ET cycles. Methods: 207 fresh ET and 194 Frozen-thawed ET (FET) cycles were included in this retrospective study. Two groups (A and B) were defined based on the EL technique used. In group A, the entire catheter was flushed with Ham's F-10 medium. The embryos were then drawn into the catheter using one air bracket. In group B, $70{\mu}L$ of air was aspirated into the syringe and the catheter was flushed using Ham's F10 medium. The medium, air, embryos, air, and finally another layer of medium were then sequentially drawn into the catheter. The main outcome measures were the pregnancy and delivery rates. Results: The groups did not differ with respect to the etiology of infertility, the source of spermatozoa, the quality of the embryos, the type of EL catheter, and the ease of transfer. The pregnancy rate was similar between two groups. In fresh ET cycles, a higher delivery rate was observed in group B than it group A (78.1% vs. 60%, p=0.1). In FET cycles, the rate of delivery was significantly higher in group B than in group A to a nonsignificant extent (88.9% vs. 58.8%, p=0.06). Conclusion: EL techniques did not have a significant impact on the delivery rate in either fresh or FET cycles.

산수유 추출방법에 따른 항산화 기능 분석 (Antioxidative Activity of Cornus officianalis Extracts Obtained by Four Different Extraction Techniques)

  • 박은비;김혜선;신소연;지인해;김지현;김성구;유병홍;김병우;곽인석;김문무;정경태
    • 생명과학회지
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    • 제22권11호
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    • pp.1507-1514
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    • 2012
  • 산수유는 다양한 생리적 기능을 가지는 것으로 보고되어 있으며 또한 최근에 관심의 증가로 산수유로부터 유효성분의 추출 방법 개발은 산업적으로 매우 중요하며, 유효성분의 존재 여부를 분석하는 것은 산업적 상품의 진위와 품질을 판정하는데 중요한 수단이 된다. 본 연구는 loganin을 지표 물질로 하여 산수유로부터 유효성분을 추출하는 여러 방법을 비교 분석하였으며, 각 방법에 따라 추출한 시료의 기능성의 차이를 항산화 작용으로 분석하였다. 산수유로부터 항산화 성분을 추출하기 위하여 네 가지 방법을 적용하였다. 추출된 유효성분 지표물질로 loganin의 함량을 HPLC로 분석하였을 때 열수를 사용한 추출 방법이 알코올 추출 또는 효소 처리 방법 보다 효율적인 결과를 보여 주었다. 각 추출 시료의 항산화 기능은 DPPH radical 소거능, hydrogen peroxide scavenging assay, reducing power 측정으로 분석하였다. 추출 방법에 따라 항산화 기능이 차이가 있었으며, 열수 추출 시료가 DPPH radical 소거능과 hydrogen peroxide scavenging 효과가 우수하였고, 효소 처리 추출 시료는 reducing power가 우수하게 나타났다. 추출 방법에 따라 시료에 함유된 성분의 차이가 항산화 기능이 차이와 연관이 있다고 생각된다. 이 연구에서 얻은 결과는 산수유는 대조군으로 사용한 vitamin C에 준하는 항산화 성분을 함유하고 있으며, 이들 성분은 열수 추출 방법으로 효율적으로 추출되었으며, 열수 추출 방법은 산업적으로 응용될 수 있음을 제시하고 있다.

Congenital Malformations in Infants of Mothers Undergoing Assisted Reproductive Technologies: A Systematic Review and Meta-analysis Study

  • Hoorsan, Hayedeh;Mirmiran, Parvin;Chaichian, Shahla;Moradi, Yousef;Hoorsan, Roza;Jesmi, Fatemeh
    • Journal of Preventive Medicine and Public Health
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    • 제50권6호
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    • pp.347-360
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    • 2017
  • Objectives: This meta-analysis aimed to evaluate congenital malformations in infants conceived by assisted reproductive techniques (ART), compared with infants conceived spontaneously. Methods: In this study, available resources searched to find relevant articles included PubMed, ScienceDirect, Scopus, Google Scholar, Cochrane, ProQuest, Iranmedex, Magiran, and Scientific Information Database. After extracting the necessary information from evaluated articles, meta-analysis on the articles' data was performed using Stata version 11.2. Results: In this study, from a total of 339 articles, extracted from the initial investigation, ultimately 30 articles were selected for meta-analysis that assessed the use of ART on the risk of congenital abnormalities and some birth complications on 5 470 181 infants (315 402 cases and 5 154 779 controls). The odds ratio (95% confidence interval [CI]) for low birth weight was 1.89 (95% CI, 1.36 to 2.62), preterm labor 1.79 (95% CI, 1.21 to 2.63), cardiac abnormalities 1.43 (95% CI, 1.27 to 1.62), central nervous system abnormalities 1.36 (95% CI, 1.10 to 1.70), urogenital system abnormalities 1.58 (95% CI, 1.28 to 1.94), musculoskeletal disorders 1.35 (95% CI, 1.12 to 1.64), and chromosomal abnormalities in infants conceived by ART was 1.14 (95% CI, 0.90 to 1.44), which were all statistically significant, except chromosomal abnormalities. Conclusions: The risk of congenital abnormalities and some birth complications were significantly higher in ART than normal conception, while chromosomal abnormalities were not; therefore, the application of ART should be selected individually for patients by detailed assessment to reduce such risks in the population.

Effects of Tween 80 on In Vitro Fermentation of Silages and Interactive Effects of Tween 80, Monensin and Exogenous Fibrolytic Enzymes on Growth Performance by Feedlot Cattle

  • Wang, Y.;McAllister, T.A.;Baah, J.;Wilde, R.;Beauchemin, K.A.;Rode, L.M.;Shelford, J.A.;Kamande, G.M.;Cheng, K.J.
    • Asian-Australasian Journal of Animal Sciences
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    • 제16권7호
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    • pp.968-978
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    • 2003
  • The effects of monensin, Tween 80 and exogenous fibrolytic enzymes on ruminal fermentation and animal performance were studied in vitro and in vivo. In Expt 1, the effects of the surfactant Tween 80 (0.2% wt/wt, DM basis) on ruminal fermentation of alfalfa, corn and orchardgrass silages were investigated using in vitro gas production techniques. Tween 80 did not affect (p>0.05) cumulative gas production at 24 h, but it reduced (p<0.05) the lag in fermentation of all three silages. With corn silage and orchardgrass silage, gas production rates and concentrations of total volatile fatty acids (VFA) were increased (p<0.05) by Tween 80; with alfalfa silage, they were reduced (p<0.05). Tween 80 increased (p<0.05) the proportion of propionate in total VFA, and reduced (p<0.05) acetate to propionate ratios (A:P) with all three silages. In Expt 2, exogenous fibrolytic enzymes (E; at 0, 37.5 or 75 g/tonne DM), monensin (M; at 0 or 25 ppm and Tween 80 (T; at 0 or 2 L/tonne DM) were added alone or in combination to backgrounding and finishing diets fed to 320 crossbred steers in a feeding trial with a $3{\times}2{\times}$2 factorial arrangement of treatments. The backgrounding and finishing diets contained barley grain and barley silage in ratios of 57.8:42.2 and 93.5:6.5 (DM basis), respectively. Added alone, none of the additives affected DM intake (p>0.1) in the backgrounding or in the finishing period, but interactive $M{\times}T$ effects were observed in the finishing period (p=0.02) and overall (p=0.04). In the finishing period, T without M tended to reduce DM intake (p=0.11), but T with M increased (p=0.05) DM intake. Monensin increased average daily gain (ADG) during backgrounding (p=0.07) and finishing (p=0.01), and this ionophore also improved overall feed efficiency (p=0.02). Warm carcass weight was increased (p<0.001) by M, but dressing percentage was reduced (p=0.07). In the backgrounding period, T increased ADG by 7% (p=0.06). Enzymes increased (p=0.07) ADG by 5 and 6% (low and high application rates, respectively) during backgrounding, but did not affect (p>0.10) ADG during finishing, or overall feed efficiency. Whereas T enhanced the positive effects of M on ADG during backgrounding (p=0.04) and overall (p=0.05), it had no impact (p>0.1) on the effects of E. Interactions between M and T suggest that the surfactant may have potential for enhancing the positive effects of monensin on beef production, but this requires further research.