• 제목/요약/키워드: monocyte isolation

검색결과 7건 처리시간 0.031초

생약으로부터 세포분화유도물질의 검색 및 분리 및 분리 (I) (Screening and Isolation of the Cell Differentiation Inducers from Medicinal Plants (I))

  • 박은정;김진웅
    • 생약학회지
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    • 제28권4호
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    • pp.225-232
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    • 1997
  • 300 extracts derived from 100 plants were tested for their potential to induce HL-60 cell differentiation using NBT assay and NSE/SE staining methods. Morphological changes from suspended to adherent state of the cells were also observed by microscopic examination. In result, 55 extracts induced cell differentiation into monocyte/macrophage lineage in the NBT and the NSE assay.

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부유밀도구배 원심분리를 이용하여 말초혈액단핵구로부터 고순도 및 고수율의 단세포 분리방법 (Isolation of Monocytes with High Purity and Yield from Peripheral Blood Mononuclear Cells by Flotation Density Gradient Centrifugation)

  • 배재호;손철훈;박유수;김동완;김선희;강치덕
    • 생명과학회지
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    • 제19권6호
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    • pp.728-734
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    • 2009
  • 본 연구에서는, PBMC로부터 고순도 및 고수율의 단세포를 분리하기 위해 Histopaque 용액의 density 및 osmolarity를 PBS와 NaCI 용액으로 1.072 g/ml 및 335 mOsm로 조절하여 간단하고 경제적인 방법인 부유밀도구배 원심분리법을 개발하였다. 위와 같이 조절된 Histopaue 용액을 사용하여 부유밀도구배 원심분리를 시행한 결과 단세포는 Top층에서 가장 많이 회수되었다. 회수된 단세포의 순도는 71.44${\sim}$82.38%의 범위를 나타내었고 평균 74.75${\pm}$3.84% 이었다. 또한, 수율은 21.02${\sim}$53.63%의 범위를 나타내어 평균 32.62${\pm}$11.16% 이었다. 이러한 방법의 부유밀도구배 원심분리에 의해 분리된 단세포를 수지상세포로 분화 유도시켜 그 형태, 표현형, 기능적인 면을 분석해 보았을 때 전형적인 수지상세포의 기능을 나타내었다. 결론적으로, PBMC로부터 단세포를 분리하기 위해 본 연구에서 사용된 부유밀도구배 원심분리 방법은 plastic adherence, elutriation 및 immunomagnetic selection보다 용이하고 경제적인 방법이다. 나아가 이러한 장점을 바탕으로 임상단계에서 수지상세포를 안전하고, 효율적으로 배양할 수 있는 closed system에도 적용이 가능할 것으로 기대된다.

옻추출물 처리에 의한 U937 세포에서의 특정 RNA 발현 양상 (Isolation and Elucidation of Specific RNAs by Treatment of Rhus verniciflua Stokes Extract to U937 Cell)

  • 정미영;오세욱
    • 한국식품과학회지
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    • 제40권5호
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    • pp.593-598
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    • 2008
  • 사람의 혈액 내의 단핵구 U937을 모델시스템으로 이용하여 옻추출물 처리에 의해 발현이 조절되는 특정 유전자를 탐색하였다. DDRT-PCR을 이용하여 옻 추출물 처리 시 발현이 감소되는 클론을 하나 얻을 수 있었으며 이를 클로닝하고 염기서열 분석을 실시하였다. 그 결과 얻어진 유전자는 H2A histone family의 member Z와 100% 유사성이 있는 것으로 나타났다. 이 단백질은 특정 조건 하에서 특정한 유전자 발현을 증가시키는 역할을 하는 것으로 보고되고 있으나, 보다 정확한 작용기작을 알아내기 위해서는 유전자 관계 파악을 위하여 향후 계속적인 연구가 필요함을 알 수 있었다.

Identification of a novel triterpene saponin from Panax ginseng seeds, pseudoginsenoside RT8, and its antiinflammatory activity

  • Rho, Taewoong;Jeong, Hyun Woo;Hong, Yong Deog;Yoon, Keejung;Cho, Jae Youl;Yoon, Kee Dong
    • Journal of Ginseng Research
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    • 제44권1호
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    • pp.145-153
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    • 2020
  • Background: Panax ginseng Meyer (Araliaceae) is a highly valued medicinal plant in Asian regions, especially in Korea, China, and Japan. Chemical and biological studies on P. ginseng have focused primarily on its roots, whereas the seeds remain poorly understood. This study explores the phytochemical and biological properties of compounds from P. ginseng seeds. Methods: P. ginseng seeds were extracted with methanol, and 16 compounds were isolated using various chromatographic methods. The chemical structures of the isolates were determined by spectroscopic data. Antiinflammatory activities were evaluated for triterpene and steroidal saponins using lipopolysaccharide-stimulated RAW264.7 macrophages and THP-1 monocyte leukemia cells. Results: Phytochemical investigation of P. ginseng seeds led to the isolation of a novel triterpene saponin, pseudoginsenoside RT8, along with 15 known compounds. Pseudoginsenoside RT8 exhibited more potent antiinflammatory activity than the other saponins, attenuating lipopolysaccharide-mediated induction of proinflammatory genes such as interleukin-1β, interleukin-6, inducible nitric oxide synthase, cyclooxygenase-2, and matrix metalloproteinase-9, and suppressed reactive oxygen species and nitric oxide generation in a dose-dependent manner. Conclusion: These findings indicate that pseudoginsenoside RT8 has a pharmaceutical potential as an antiinflammatory agent and that P. ginseng seeds are a good natural source for discovering novel bioactive molecules.

포도상구균 장내 C 형 변이독소 (SEC mutant)의 면역원성에 대한 연구 (Immunogenicity of staphylococcal enterotoxin C mutant antigen in mice and dairy cows)

  • 장병선;주이석;문진산;서근석;양수진;김소현;박용호
    • 대한수의학회지
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    • 제41권2호
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    • pp.177-188
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    • 2001
  • Mastitis is one of the most significant cause of economic loss to the dairy industry. Especially, Staphylococcus aureus is a major contagious mastitis-causing pathogen in dairy cattle. Because of its high transmission rate and resistance to antibiotic therapy, staphylococcal mastitis presents a constant threat to the dairy industry. Staphylococcal enterotoxin C(SEC) produced by S aureus has been known as one of superantigens which are able to stimulate a large proportion of T lymphocytes independently of their antigenic specificity. In this experiment, we have conducted preliminary studies with mice and lactating cows to evaluate the immunogenicity and safety of the experimental vaccine consists of SEC mutant antigen on controlling the bovine mastitis associated with S aureus infections. The average value of somatic cell counts in quarter milk, isolation rate of S aureus were consistently decreased in SEC-SER vaccinated groups, whereas antibody titers were highly increased in SEC-SER vaccinated groups. Peripheral blood were also collected from the lactating cows to determine the proportion of leukocyte subpopulation associated with humoral immunity(HI) and cell mediated immunity(CMI). Proportion of leukocyte subpopulation expressing $BoCD2^+$(total T lymphocyte), $BoCD4^+$(T helper cell), $BoCD8^+$(T cytotoxic/suppressor cell) and NonT/NonB lymphocyte which are involved in CMI in SEC-SER vaccinated groups were decreased for the initial stage after first vaccination and then increased from ten weeks after first vaccination maintaining elevated level till 14 weeks after vaccination. In contrast, proportion of monocyte, MHC class II and B lymphocyte which are associated with the production of primary immune response in SEC-SER vaccinated groups were increased for the initial period and then decreased from ten weeks after first vaccination. We present evidence that vaccination of SEC-SER mutant antigen in lactating cows induced a significant proliferation of bovine T lymphocytes. These results suggest that SEC-SER mutant antigen used in this experiment might be one of potential immunogen in developing innovative vaccine against bovine IMI associated with S aureus. Additional challenge trials should be carried out to evaluate substantial protection against S aureus under the commercial farm conditions.

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만성육아종질환 환자 단핵구에서 TNF-α 자극에 의한 IL-8과 IL-10의 발현 양상 (TNF-α stimulated IL-8 and IL-10 expression in monocytes from patients with chronic granulomatous disease)

  • 신경수
    • Clinical and Experimental Pediatrics
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    • 제51권10호
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    • pp.1096-1101
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    • 2008
  • 목적: 만성 육아종 질환은 백혈구 내에서 반응성 산소 대사물을 생성하는 효소인 NADPH oxidase의 유전적인 결함으로 인하여 반복적이고 치명적인 세균 감염과 진균 감염이 발생되는 유전성 면역 결핍 질환이다. 만성적인 염증 상태의 원인과 염증성 육아종 형성의 기전은 잘 알려져 있지 않으나 만성육아종질환의 염증 조절 부전과 관련이 있을 것으로 생각된다. 본 연구는 만성육아종질환 환자의 단핵구에서 $TNF-{\alpha}$의 자극으로 발현되는 IL-8과 IL-10을 측정하여 염증성 시토카인과 항염증성 시토카인의 발현 양상과 만성육아종질환 환자의 만성적인 염증 상태와의 관련성을 알아보고자 하였다. 방법: 만성 육아종 질환 환자 8명과 건강 대조군 10명의 말초 혈액에서 magnet cell isolation kit를 이용하여 단핵구를 분리하고, $TNF-{\alpha}$ (10 ng/mL)로 6시간 동안 자극한 후 실시간 정량적 중합효소 연쇄반응과 ELISA법으로 IL-8과 IL-10의 발현 정도를 측정하였고, 건강 대조군의 단핵구에 항산화제를 전처리한 후 $TNF-{\alpha}$로 자극하여 IL-8과 IL-10의 발현 정도를 측정하였다. 결 과: 만성육아종질환 환자에서 $TNF-{\alpha}$ 자극에 의한 IL-8의 발현 양상은 건강 대조군과 달리 최고 발현치가 60분 정도 늦게 관찰되었고, IL-8의 발현도 자극 시간이 경과하여도 지속되었다. 그리고 건강 대조군의 단핵구에 항산화제를 처리하여 만성육아종질환과 유사한 조건하에서 IL-8을 측정하였을 때 항산화제를 처리한 군에서 처리하지 않은 대조군에 비하여 IL-8의 발현이 증가하였고, catalase와 apocynin을 처리한 군에서 DPI를 처리한 군보다 IL-8의 발현이 증가하였다. $TNF-{\alpha}$ 자극에 의한 IL-10의 발현도 IL-8의 발현 양상과 유사하게 지연 반응과 과발현이 관찰되었다. 그러나 건강 대조군의 단핵구에 항산화제를 처리하였을 때의 IL-10 발현은 처리하지 않은 군과 큰 차이를 보이지 않았다. 결 론: 만성육아종질환 환자의 단핵구에서 $TNF-{\alpha}$ 자극에 의하여 염증성 시토카인인 IL-8과 항염증성 시토카인인 IL-10의 지연 반응과 과발현이 동시에 관찰되었으며, 건강 대조군의 단핵구에 항산화제를 처리하였을 때 IL-8의 발현 증가가 관찰되었다. 이는 만성육아종질환의 만성 염증 상태가 시토카인의 생성 조절의 부전과 연관이 있고, NADPH oxidase의 결함에 의한 반응성 산소대사물의 생성 부전과도 연관되어 있음을 알 수 있다.

서울지역에서 문제되고 있는 강아지의 급성설사증에 관한 병인학적 연구 (Etiologic Survey on the Acute Diarrhea of Puppies in Seoul)

  • 한홍율;박희명;이진희;오태호;정순욱;윤신근;박철만
    • 한국임상수의학회지
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    • 제10권2호
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    • pp.171-184
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    • 1993
  • In Seoul area, there are so many kennel clubs, veterinary hospital, pet establishment and breeding confers that various problems have occurred. They are crowed pet houses, poor sanitation, stress to puppies, sudden environmental changes to puppies and unvaccination against parvovirus, canine distemper virus, parainfluenza virus, infectious hepatitis caused by Adenovirus type I and Leptospira. Several studies were made to survey the infectious agents involved in acute diarrhea of poppies in Seoul are, such as history taking, physical examination, complete blood count and serum chemistry, histopathological finding, bacterial isolation and identification, and hemagglutination test in feces and hemagglutination inhibition test in serum against parvovirus, respectively. The results obtained are summarized as followed. 1. The percent of PCV (30.5$\pm$5.6) and concentration of Total protein(5.0$\pm$0.8) resulted from statistical analysis are significantly lower than normal values (p<0.05), respectively. In addition, fibrinogen (505$\pm$326) was significantly higher an normal value (p<0.001), Band neutrophil (22.9$\pm$12.7) showed signifiant difference (P<0.01). decreased monocyte (3.2$\pm$2.1) and eosinophil (0.7$\pm$0.8) values appeared statistically significant (p<0.001), lymphocyte (16.7$\pm$9), as well. 2. The concentrations of calcium .(8.0$\pm$2.8), glucose (40.1$\pm$31.4), and albumin (2.0$\pm$0.39) were lower than normal values (p<0.01, p<0.01, p<0.001), respectively. Also Inorganic phosphate (7.1$\pm$2.4), pH (p<7.9f:0.2), and Blood Urea Nitrogen (40.41=37.1) were significantly higher than normal values (p<0.001, p(0.001, p<0.05), respectively. 3. Simple and mixed infections occupied 18% and 82% in the distribution of causes in puppies with acute diarrhea, respectively. 4. As puppies got older, incidence of acute diarrhea caused by Staphylococcus aureus was decreased to 13% and infection of canine distemper virus was increased to 53%, but E coli and canine parvovirus always showed high frequency of outbreak in the body weight ranged from 35g to 7.8Kg. 5. As showed in table 5, infections of E coli and Canine Parvovirus showed high outbreak regardless of the age which is classified into three stages, 35~50 day, 60 day and 75~day to 1 year, canine distemper virus appeared increased, but in case of Staphylococcus aureus, visa versa. 6. In comparison wi methods for the laboratory diagnosis diagnosis parvovirus, Hemagglutination test showed positive reaction in 25% and mean serum antibody titer measured by Hemagglutination inhibition test showed 2779 (n=20). In addition, positive reaction was 90% (18/20). 7. In histopathological studies, enteritic and pneumonic lesions were indicated in 53.7% and 39.5% of samples, respectively. 8. Etiologic diagnosis based on the history taking, clinical signs and histopathological findings in puppies with acute diarrhea and vomiting indicated that 50% of puppies were infected by canine parvovirus and distemper virus. 9. In the parasitological examination made by simple flotation with saturated zinc sulfate tour parasites found were Isospora canis, Toxocara canis, Ancylostoma spp and Toxocara spp. Isospora canis and Toxocara canis were more frequently found among those parasites of diarrhoeic causes in puppies ranged from 35 days to 1 year. Their infestation rates were 15% and 13% respectively.

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