• 제목/요약/키워드: co-culture system

검색결과 474건 처리시간 0.026초

HeLa 세포의 Spheroid에 대한 방사선과 Platinum 유사체의 치사 효과 (Lethal Effects of Radiation and Platinum Analogues on Multicellular Spheroids of HeLa Cells)

  • 홍성언
    • Radiation Oncology Journal
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    • 제7권2호
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    • pp.149-156
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    • 1989
  • HeLa 세포의 spheroid를 배 양시켜 cis-platinum과 carboplatin으로 처리한 후 그 반응을 세포의 생존분획으로 분석하였다. 체외실험 model인 spheroid를 사용하여 platinum유사체의 약효와 방사선 감수성을 평가하고 약제에 대한 단층 세포와 spheroid의 감수성 차이를 세포-생존곡선에서 규명하기 위하여 본 실험을 시행하였다. Cis-platinum 농도-곡선에서 spheroid의 $Cq=3.4{\mu}M$이고 $Co=1.2{\mu}M$이었다. 이 것은 단층세포에 비하여 Co는 큰 변화가 없으나 Cq가 증가되어 cis-platinum이 저산소층 세포보다 활동적으로 분화하는 표면세포에 주로 작용하였으며, 반대로 carboplatin의 효과는 spheroid에 대한 $(Co=15.0{\mu}M)$ 감수성이 단층세포$(Co=32.5{\mu}M)$에 비하여 크게 증가되어, spheroid의 심층 세포에 주로 작용하였다. 방사선과 carboplation의 병용효과를 세포 생존분획이 0.01 수준에서 isobologram으로 분석한 결과 상호작용으로 supra-additive 효과를 나타내었다. 따라서, carboplatin은 cis-platinum에 비하여 신장과 위장에 대한 독성작용이 적고, 방사선과 병용함으로써 향후 더욱 효과적 인 종양 치료에 중요한 역할을 할 것으로 기대한다.

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Cyclic AMP대사가 Toxoplasma gondii의 체외 배양에 미치는 영향 (The effect of cyclic AMP on the growth of Toxoplasma gondii in vitro)

  • 최원영;남호우
    • Parasites, Hosts and Diseases
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    • 제28권2호
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    • pp.71-78
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    • 1990
  • 세포내 cAMP의 농도 변화가 Toxoplasma의 성장 및 증식에 미치는 영향을 검토하기 위하여 끽접 cAMP와 cAMP의 analogue를 첨가하거나 간접적으로 세포내 cAMP의 합성이나 분해를 담당한 효소들의 활성을 변화시 킴으로써 CAMP의 농도를 조절하여 그 효과를 측정하였다. HL-60세포를 숙주 세포로 사용하여 동수의 Toxoplasma를 첨가하여 배양하였으며, 처리 효과는 배양 세포계에서 Toxoplnsma에만 특이하게 표지되는 $^3H-uracil$과 Toxoplasma및 HL-60세포에 공통으로 표지되는 $^3H-thymidine$을 각각 매 12시간마다 2시간씩 배양하여 그 표지량을 측정하여 비교 분석하였다. 직접 cAMP와 cAMP의 analogue인 dbcAMP 및 br-cAMP를 첨가하였을 때, 각각 특이한 농도에서, 즉 1 mM, 0.5~5mM 및 0.1mM에서 Toxoplasma의 성장 및 증식을 향상시켰다. 이때, 성장 및 증식을 유도하는 시기 및 그 증가도에서도 차이가 있는 것으로 나타났다. 이 결과들은 세포내 cAMP의 농도를 증가시키도록 cAMP의 합성 및 분해에 관여하는 효소들의 황성을 변화 시켰을 때에도 같은 양상으로 나타났다. cAMP의 합성 효소인 adenylate cyclase의 활성제인 pNHppG, cAMP의 분해 효소인 cAMP phosphodiesterase의 활성제인 A23187과 imidazole 및 억제제인 IBMX, compound 48/80 및 theophylline을 각각 처리하였는데, 세포내 cAMP의 농도가 증가되었을 대에는 Toxoplasmn의 성장 및 증식 을 향상시켰으나, cAMP의 농도를 감소시켰을 때에는 억제하였다. 이 때 배양계에 독성을 일으키는 농도 이하에서 농도 의존성의 경향을 보였으며, 유도 시기 및 그 증가도에는 차이가 있었다. 따라서 세포 내에서 향상된 수준의 cAMP가 어떤 기전을 활성화시키며, 그 결과 세포질에서의 Toxcplasma의 성장 및 증식을 자극하게 된다는 것을 시사하였다.

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THE STUDY ON TISSUE CULTURED WILD MOUNTAIN GINSENG(Panax Ginseng C.A. Meyer) ADVENTITIOUS ROOTS EXTRACT AS A COSMETIC INGREDIENT

  • Jung, Eun-Joo;Park, Jong-Wan;Kim, Joong-Hoi;Paek, Kee-Yoeup
    • 대한화장품학회:학술대회논문집
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    • 대한화장품학회 2003년도 IFSCC Conference Proceeding Book I
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    • pp.611-616
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    • 2003
  • Korean ginseng(Panax Ginseng C.A. Meyer) known as a oriental miracle drug is an important medicinal plant. Ginseng has been used for geriatric, tonic, stomachic, and aphrodisiac treatments for thousands years. Also, it is an antibiotic and has therapeutic properties against stress and cancer. Ginseng is widely distributed all over the world. Among them, Korean mountain ginseng has the most valuable effect on pharmaceuticals. The roots of mountain ginseng contained several kinds of ginsenosides that have many active functions for the human body. However, the study of mountain ginseng has a limit because the mountain ginseng is very expensive and rare. So, we artificially cultured mountain ginseng adventitious roots using the bioreactor culture system. We induced callus from original mountain ginseng, directly dug up in mountain and aged about one hundred ten years. Separated adventitious roots were precultured in 500ml conical flasks and then, transferred in 20L bioreactors. The adventitious roots of mountain ginseng were harvested after culturing for 40days, dried and then, extracted with several solvents. In this study, we investigated the whitening effect, anti-wrinkle effect and the safety of tissue cultured adventitious roots extract of mountain ginseng in order to identify the merit as a cosmetic ingredient. Particularly, extract of mountain ginseng adventitious roots showed whitening and anti-wrinkle effects. The inhibitory effect of this extract on the melanogenesis was examined using B-16 melanoma cell. When B-16 melanoma cells were cultured with adventitious root extract, there was a dramatically decrease in melanin contents of 8-16 melanoma cell. And we identified this extract inhibited Dopa auto-oxidation significantly. Also, when transformed mouse fibroblast L929 cells were treated with this extract, there was a significant increase in collagen synthesis. The results show significant inhibited melanization and wrinkle without inhibiting cell viability.

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Intraventricular Antimicrobial Therapy for Intractable Ventriculitis: Two Case Reports

  • Lee, Ji Weon;Yoon, Yoonsun;Kim, Sang-Dae;Kim, Yun-Kyung
    • Pediatric Infection and Vaccine
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    • 제29권1호
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    • pp.46-53
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    • 2022
  • 뇌실염은 일반적으로 정맥 내 항생제를 통하여 치료하나, 불응성 뇌실염의 치료는 정맥 내 항생제 치료와 뇌실 내 항생제 치료의 병행이 요구되기도 한다. 이론적으로 항생제의 뇌실 내 투여는 정맥 내 단독 투여보다 뇌척수액에서 더 높은 항생제 농도에 도달할 수 있게 한다. 본 증례 보고는 기존의 전신 항생제 치료에 불응하는 폐렴 간균과 메티실린 내성 표피 포도상 구균에 각각 뇌실 내 겐타마이신과 반코마이신 투여를 통하여 뇌실염을 치료한 2례로서 이후에도 주요 합병증 등이 없어 이를 보고하는 바이다.

부자와 육계 병용투여 시 파골세포 분화 억제에 미치는 영향 (Effect of Co-administration of Aconiti Lateralis Preparata Radix and Cinnamomi Cortex on Osteoclast Differentiation)

  • 정기은;김정영;김지훈;한상용;김윤경
    • 대한본초학회지
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    • 제29권2호
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    • pp.61-67
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    • 2014
  • Objectives : Aconiti Lateralis Preparata Radix (Aconitum Carmichaeli, AC) and Cinnamomi Cortex (Cinnamomi Cortex, CC) have been treated to elderly for kidney yang enhancement in Korean traditional medicine. In this study, the effects of water extract of AC and CC on RANKL (Receptor Activator for Nuclear Factor ${\kappa}B$ Ligand)-induced osteoclast differentiation were evaluated in culture system. Methods : MTT assay was used to evaluate the potential cytotoxicity of AC and CC extracts in bone macrophage marrows (BMMs) stimulated with M-CSF. TRAP (tartrate-resistant acid phosphatase) staining and TRAP activity were performed to know the inhibitory effect on osteoclast differentiation. The protein expression levels of nuclear factors such as activated T cell(NFAT)c1, c-Fos, MAPKs and ${\beta}$-actin in cell lysates treated with AC and CC extracts were analysed by western blotting. Results : AC, CC extracts and their co-administration inhibited significantly RANKL-induced osteoclast differentiation in BMMs in a dose dependent manner without toxicity. Each AC and CC extracts inhibited the phosphorylation of p38. Also, AC and CC extracts, respectively, inhibited the protein expression of c-Fos and NFATc1 more than Co-administration of AC and CC even if all treatments did. It was observed that RANKL-induced degradation of I-${\kappa}B$ is significantly suppressed by all treatments. Conclusions : Taken together, It was concluded that AC and CC have beneficial effect on osteoporosis by inhibition of osteoclast differentiation. Thus, Atractylodis AC and CC could be a treatment option for osteoporosis.

Effect of Thyroid hormone on Lipogenesis in Rat White and Brown Adipocytes Culture System

  • Kim, Yangha -Moon
    • Preventive Nutrition and Food Science
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    • 제3권4호
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    • pp.362-367
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    • 1998
  • Thyroid hormone(T3) stimulates hepatic lipogenesis by increasing expression of genes, indluding acetyl-CoA carboxylase and fatty acid synthase. S14 protein, which is thougth to be involved in lipid metabolism , appears to respond in parallel . Effect of T3 on lipogenesis in white and brown adipose tissue are less clear, and may be complicated by indirect effects of the hormone. We developed an adipocytes system where the indirect effects of thyroid hormone are abolished and direct effects of T3 on lipogenesis could be tested. Fat accumulation was mesured by Oil-Red O staining. Insulin clearly enhanced fat accumulation by 2-fold . Isobutylemethylxanthie(IBMX) apeared to inhibit insulin -stimulated fat accumulation. Dexamethasone increased insulin-stimulatedfat accumulation about 1.3-fold. confluent adipocytes were cultured in serum-free medium or medium containing 10% fetal calf serum or 10% fetal calf serum stripped of thyroid hormone and lipogenesis, assessed by the incorporation of 3H2O , was measured. Medium without serum or supplemented with T3-depleted serum did not amplify the stimulatory effect of T3 on lipogenesis compared to medium containing 10% fetal calf seru. Dexamethasone alone led to a decrease inlopogenesis of about 50 % in white adipocytes and 25% in brown adipocytes. However, dexamethasone amplified the lipogenic respnse to T3 by about 30% in whit eadipocytes and 60% in brown adipocytes. T3(1$\mu$M) stimulated lipogenesis and acetyl-CoA carboxylase and fatty acid syntase mRNA levels up to 2 -fold in both types of adipocytes. It seems that these adipocytes systems are as useful model to study the effects of hormones on lipogenic gene expression as well as lipogenesis.

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Efficient transgene expression system using a cumate-inducible promoter and Cre-loxP recombination in avian cells

  • Park, Tae Sub;Kim, Si Won;Lee, Jeong Hyo
    • Asian-Australasian Journal of Animal Sciences
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    • 제30권6호
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    • pp.886-892
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    • 2017
  • Objective: Transgenic technology is widely used for industrial applications and basic research. Systems that allow for genetic modification play a crucial role in biotechnology for a number of purposes, including the functional analysis of specific genes and the production of exogenous proteins. In this study, we examined and verified the cumate-inducible transgene expression system in chicken DF1 and quail QM7 cells, as well as loxP element-mediated transgene recombination using Cre recombinase in DF1 cells. Methods: After stable transfer of the transgene with piggyBac transposon and transposase, transgene expression was induced by an appropriate concentration of cumate. Additionally, we showed that the transgene can be replaced with additional transgenes by co-transfection with the Cre recombinase expression vector. Results: In the cumate-GFP DF1 and QM7 cells, green fluorescent protein (GFP) expression was repressed in the off state in the absence of cumate, and the GFP transgene expression was successfully induced in the presence of cumate. In the cumate-MyoD DF1 cells, MyoD transgene expression was induced by cumate, and the genes controlled by MyoD were upregulated according to the number of days in culture. Additionally, for the translocation experiments, a stable enhanced green fluorescent protein (eGFP)-expressing DF1 cell line transfected with the loxP66-eGFP-loxP71 vector was established, and DsRed-positive and eGFP-negative cells were observed after 14 days of co-transfection with the DsRed transgene and Cre recombinase indicating that the eGFP transgene was excised, and the DsRed transgene was replaced by Cre recombination. Conclusion: Transgene induction or replacement cassette systems in avian cells can be applied in functional genomics studies of specific genes and adapted further for efficient generation of transgenic poultry to modulate target gene expression.

ICT 기반 스마트 재배시스템을 이용한 고온기 표고재배 환경변화 조사 (Research of environmental condition using ICT-based smart cultivation system during high temperature period)

  • 김인엽;권형일;황인호;이원호;노종현;최선규;고한규;구창덕
    • 한국버섯학회지
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    • 제16권2호
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    • pp.79-85
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    • 2018
  • ICT 기반 스마트 표고재배시설을 활용하여 표고재배의 과정을 후숙관리, 발이관리, 생육관리 및 휴양관리 4과정으로 나누어 6월부터 10월까지 시험을 진행하였다. 여름철 재배사 외부 $20{\sim}35^{\circ}C$의 고온환경에서도 재배사 내부의 환경은 설정한 방향으로 제어되었다. 여름철 대표적인 재배용 품종 '산조701호'와 신품종 '산조717호'를 이용한 재배시험 결과 '산조701호'의 경우 353.7 g/봉, '산조717호'는 270.4 g/봉으로 '산조701호'가 높은 버섯생산성을 나타내었다. ICT 기반으로한 스마트 표고재배시설을 활용한 표고재배는 재배과정별 환경조건 설정으로 재배가 어려운 고온의 환경에서도 편리하게 재배환경 관리가 가능하였으며, 재배품종에 적합한 환경제어 범위를 연구한다면 버섯생산 안정화 및 생산성 향상에 크게 기여할 것으로 판단되었다.

Xylitol Down-Regulates $1{\alpha},25$-Dihydroxy Vitamin D3-induced Osteoclastogenesis via in Part the Inhibition of RANKL Expression in Osteoblasts

  • Ohk, Seung-Ho;Jeong, Hyunjoo;Kim, Jong-Pill;Yoo, Yun-Jung;Seo, Jeong-Taeg;Shin, Dong-Min;Lee, Syng-Ill
    • International Journal of Oral Biology
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    • 제38권3호
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    • pp.127-134
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    • 2013
  • Xylitol is a sugar alcohol with a variety of functions including bactericidal and anticariogenic effects. However, the cellular mechanisms underlying the role of xylitol in bone metabolism are not yet clarified. In our present study, we exploited the physiological role of xylitol on osteoclast differentiation in a co-culture system of osteoblastic and RAW 264.7 cells. Xylitol treatment of these co-cultures reduced the number of tartrate-resistant acid phosphatase (TRAP)-positive multinucleated cells induced by 10 nM $1{\alpha},25(OH)_2D_3$ in a dose-dependent manner. A cell viability test revealed no marked cellular damage by up to 100 mM of xylitol. Exposure of osteoblastic cells to xylitol decreased RANKL, but not OPG, mRNA expression in the presence of $10^{-8}M$ $1{\alpha},25(OH)_2D_3$ in a dose-dependent manner. Furthermore, bone resorption activity, assessed on bone slices in the coculture system, was found to be dramatically decreased with increasing xylitol concentrations. RANKL and OPG proteins were assayed by ELISA and the soluble RANKL (sRANKL) concentration was decreased with an increased xylitol concentration. In contrast, OPG was unaltered by any xylitol concentration in this assay. These results indicate that xylitol inhibits $1{\alpha},25(OH)_2D_3$-induced osteoclastogenesis by reducing the sRANKL/OPG expression ratio in osteoblastic cells.

호염기성 미세조류 Arthrospira platensis의 폐수처리 적용을 위한 종특이성 평가 (Species Specificity Evaluation for Wastewater Treatment Application of Alkaliphilic Microalgae Arthrospira platensis)

  • 이수현;허재희;황선진
    • 한국물환경학회지
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    • 제38권6호
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    • pp.282-291
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    • 2022
  • Since the efficiency of wastewater treatment using microalgae differs depending on the metabolic characteristics of the species, it is important to understand the characteristics of target algae prior to the application in wastewater treatment. In this study, for the application of Arthrospira platensis to wastewater treatment, which is a filamentous alkaliphilic cyanobacteria, basic species specificity was identified and the possibility of application to wastewater treatment was investigated. As a result of the species specificity investigation, the specific growth rate between pH 7.0 and 11.0 showed the highest value near pH 9 at 0.25/day. The reason for the relatively low growth(0.08/day) at pH 11 was thought to be the CA(carbonic anhydrase) enzyme that is involved in carbon fixation during photosynthesis has the highest activity at pH 8.0 to 9.0, and at pH 11, CA activity was relatively low. In addition, A. platensis showed optimal growth at 400 PPFD(photosynthetic photon flux density) and 30℃, and this means that cyanobacteria such as A. platensis have a larger number of PS-I(photosystem I) than that of PS-II(photosystem II). It was speculated that it was because higher light intensity and temperature were required to sufficiently generate electrons to transfer to PS-I. Regarding the applicability of A. platensis, it was suggested that if a system using the synergistic effect of co-culture of A. platensis and bacteria was developed, a more efficient system would be possible. And different from single cocci, filamentous A. platensis expected to have a positive impact on harvesting, which is very important in the latter part of the wastewater treatment process.