• 제목/요약/키워드: colony stimulating activity

검색결과 70건 처리시간 0.032초

Enhanced production of hGM-CSF by temperature shifting in transgenic Nicotiana tabacum cell suspension cultures

  • Kim, Yong-Hoon;Lee, Sang-Yoon;Kim, Dong-Il
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2003년도 생물공학의 동향(XIII)
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    • pp.329-333
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    • 2003
  • 본 연구에서는 형질전환된 N. tabacum 배양에 있어서 배양 중반 저온으로의 변환이 세포에 미치는 영향과 hGM-CSF의 생산성 변화를 관찰하였다. 배양 중반 저온으로의 변환은 DCW의 증가와 세포크기의 감소를 보였다. Ascorbic acid의 첨가는 배양초기 세포 생존율의 감소를 완화시켰으며, 배양 중반 저온으로의 변환은 약간의 세포생존율 감소를 보였다. 저온으로의 변환, 저온 배양에서의 betaine 첨가, ascorbic acid 첨가 모두 배양 후반 세포 lysis 억제에 효과가 있었다. 배양 중반 저온으로의 변환시 배지내 단백질 분해 효소의 활성을 측정한 결과, 대조구 세포에 비해 낮은 단백질 분해 효소 활성을 나타내었다. 그로인해 배양 중반 이후 단백질 분해 효소에 의한 급격한 hGM-CSF 분해를 감소시킴으로써 상대적으로 대조구 세포에 비해 높은 hGM-CSF 생산성을 유지시켰다. Ascorbic acid를 첨가한 후 배양 도중 betaine(1 mM)을 첨가하여 저온으로 온도를 변환시, hGM-CSF의 생산성 대조구 세포에 비하여 최대 2.1배 까지 높게 유지시켰다.

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Immobilization of transgenic Nicotiana tabacum cell suspensions for the continuous production of hGM-CSF

  • Roh, Yun-Sook;Lee, Sang-Yoon;Kim, Dong-Il
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2003년도 생물공학의 동향(XIII)
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    • pp.341-345
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    • 2003
  • 형질전환된 담배세포 배양을 이용한 hGM-CSF 생산에 있어 polyurethane foam 내 고정화법을 이용하여 연속배양의 가능성을 확인하였고 spinner flask에서의 배양에 따른 영향을 알아보았다. 16일 동안 3번의 배지교환에도 세포의 활성이 유지되어 hGM-CSF 생산량은 계속적으로 증가하였다. 온도와 교반속도를 동일하게 하였을 때 spinner flask에서의 hGM-CSF 생산량은 17.3 ${\mu}g/L$으로 100-mL flask의 9.8 ${\mu}g/L$보다 증가하였다. 따라서 최적의 배지교환 속도와 양을 결정하여 spinner flask에서 연속배양을 실시할 경우 세포 재사용이라는 경쟁력과 함께 높은 hGM-CSF 생산량이 얻어질 것으로 예상된다.

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In vitro and In vivo Effects of Gelidium amansii on Intestinal Immune System

  • Jun, Woo-Jin;Kim, Se-Han;Lee, Dae-Hee;Chun, Jin-Woong;Sim, Sang-In;Lee, Kwang-Won;Cho, Hong-Yon;Hong, Bum-Shik
    • Food Science and Biotechnology
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    • 제14권1호
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    • pp.147-151
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    • 2005
  • Purified compound with intestinal immune system-modulating properties, GWE-2c, was isolated from methanol extract of Gelidium amansii by sequential procedures with silica gel column, LH-20 Sephadex gel column, and thin-layer chromatographies. In the presence of GWE-2c, strong immunoactivity in Peyers patch cell-mediated bone marrow cells was observed in vitro. In vivo intestinal immune-modulating activity was also enhanced by crude phenolic compound (GWE) of G. amansii in a dose-dependent manner. Investigation of production of several cytokines in Peyer's patch cells upon stimulation with GWE in vivo revealed the levels of granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin (IL)-6 increased. Results suggest that the phenolic compound from G. amansii represents immunopotentiator and biological response modifier at in vitro and in vivo levels.

Stability Enhancement of hGM-CSF in Transgenic Nicotiana tabacum Suspension Cell Cultures

  • Lee, Sang-Yoon;Cho, Jong-Moon;Kim, Dong-Il
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제8권3호
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    • pp.187-191
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    • 2003
  • Proteolytic enzymes existing in plant cell cultured media are the major reason for the loss of secreted human granulocyte-macrophage colony-stimulating factor (hGM-CSF). The addition of pepstatin, aprotinin and PMSF relatively decreased the proteolytic degradation of hGM-CSF in a conditioned medium, but sufficient prevention against the proteolytic activity could not be obtained with chemical protease inhibitors. Gelatin, as a competitive substrate for protease, showed a stabilizing effect in a conditioned medium. Compared to the initial hGM-CSF concentration in a conditioned medium. with 10 g/L of gelatin, 68% of the hGM-CSF remained after 5 days. In a cell culture experiment, 5 g/L of gelatin significantly stimulated the hGM-CSF production and accumulation in culture media, with no growth inhibition. compared to the controls (4.72 $\mu\textrm{g}$/L), the extracellular hGM-CSF level could be increased to 39.78 $\mu\textrm{g}$/L with the addition of 5 g/L of gelatin.

Dendritic cells resist to disulfiram-induced cytotoxicity, but reduced interleukin-12/23(p40) production

  • Haebeen Jung;Hong-Gu Joo
    • The Korean Journal of Physiology and Pharmacology
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    • 제27권5호
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    • pp.471-479
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    • 2023
  • Disulfiram (DSF), a medication for alcoholism, has recently been used as a repurposing drug owing to its anticancer effects. Despite the crucial role of dendritic cells (DCs) in immune homeostasis and cancer therapy, the effects of DSF on the survival and function of DCs have not yet been studied. Therefore, we treated bone marrow-derived DCs with DSF and lipopolysaccharide (LPS) and performed various analyses. DCs are resistant to DSF and less cytotoxic than bone marrow cells and spleen cells. The viability and metabolic activity of DCs hardly decreased after treatment with DSF in the absence or presence of LPS. DSF did not alter the expression of surface markers (MHC II, CD86, CD40, and CD54), antigen uptake capability, or the antigen-presenting ability of LPS-treated DCs. DSF decreased the production of interleukin (IL)-12/23 (p40), but not IL-6 or tumor necrosis factor-α, in LPS-treated DCs. We considered the granulocyte-macrophage colony-stimulating factor (GM-CSF) as a factor to make DCs resistant to DSF-induced cytotoxicity. The resistance of DCs to DSF decreased when GM-CSF was not given or its signaling was inhibited. Also, GM-CSF upregulated the expression of a transcription factor XBP-1 which is essential for DCs' survival. This study demonstrated for the first time that DSF did not alter the function of DCs, had low cytotoxicity, and induced differential cytokine production.

Characterization of Aspergillus niger Mutants Deficient of a Protease

  • Chung, Hea-Jong;Park, Seung-Moon;Kim, Dae-Hyuk
    • Mycobiology
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    • 제30권3호
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    • pp.160-165
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    • 2002
  • Aspergillus niger has been used as a host to express many heterologous proteins. It has been known that the presence of an- abundant protease is a limiting factor to express a heterologous protein. The protease deficient mutant of A. niger was obtained using UV-irradiation. A total of $1{\times}10^5$ spores were irradiated with $10{\sim}20%$ survival dose of UV, 600 $J/m^2$ at 280 nm, and the resulting spores were screened on the casein-gelatin plates. Ten putative protease deficient mutants showing the reduced halo area around colonies were further analyzed to differentiate the protease deficient mutant from other mutant types. Among ten putative mutants, seven mutants showed significant growth defect on nutrient rich medium and two mutants appeared to be the secretory mutants, which resulted in the impaired secretion of extracellular proteins including proteases. A mutant $pro^--20$ showed reduced halo zone without any notable changes in growth rate. In addition, the starchdegrading and glucose oxidase activities in the culture filtrate of $pro^--20$ mutant showed the similar range as that of the parental strain, which suggested that the $pro^--20$ mutant ought to be the protease deficient mutant rather than a secretory mutant. The reduced proteolytic activity of the $pro^--20$ was demonstrated using SDS-fibrin zymography gel. The reduced extracellular proteolysis was quantified by casein degradation assay and, comparing with the parental strain, less than 30% residual extracellular protease activity was detected in the culture filtrate of the $pro^--20$ mutant. The bio-activity of an exogenously supplemented hGM-CSF(human Granulocyte-Macrophage Colony Stimulating Factor) in the culture filtrate of $pro^--20$ mutant was detected until eight times more diluted preparations than that of the parental strain.

Acacia ferruginea Inhibits Tumor Progression by Regulating Inflammatory Mediators-(TNF-α, iNOS, COX-2, IL-1β, IL-6, IFN-γ, IL-2, GM-CSF) and Pro-Angiogenic Growth Factor-VEGF

  • Sakthivel, Kunnathur Murugesan;Guruvayoorappan, Chandrasekaran
    • Asian Pacific Journal of Cancer Prevention
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    • 제14권6호
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    • pp.3909-3919
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    • 2013
  • The aim of the present investigation was to evaluate the effect of A ferruginea extract on Dalton's lymphoma ascites (DLA) induced tumours in BALB/c mice. Experimental animals received A ferruginea extract (10 mg/kg.b.wt) intraperitoneally for 14 consecutive days after DLA tumor challenge. Treatment with extract significantly increased the life span, total white blood cell (WBC) count and haemoglobin (Hb) content and decreased the level of serum aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), gamma glutamyl transferase (${\gamma}$-GT) and nitric oxide (NO) in DLA bearing ascites tumor models. In addition, administration of extract significantly decreased the tumour volume and body weight in a DLA bearing solid tumor model. The levels of pro-inflammatory cytokines such as tumor necrosis factor-alpha (TNF-${\alpha}$), interleukin-1 beta (IL-$1{\beta}$), interleukin-6 (IL-6) and granulocyte monocyte-colony stimulating factor (GM-CSF), as well as pro-angiogenic growth factors such as vascular endothelial growth factor (VEGF) and inducible nitric oxide synthase (iNOS) were elevated in solid tumour controls, but significantly reduced by A ferruginea administration. On the other hand, the extract stimulated the production of interleukin-2 (IL-2) and interferon-gamma (IFN-${\gamma}$) in animals with DLA induced solid tumours. Increase in $CD4^+$ T-cell population suggested strong immunostimulant activity for this extract. GC/MS and LC/MS analysis showed quinone, quinoline, imidazolidine, pyrrolidine, cyclopentenone, thiazole, pyrazole, catechin and coumarin derivatives as major compounds present in the A ferruginea methanolic extract. Thus, the outcome of the present study suggests that A ferruginea extract has immunomodulatory and tumor inhibitory activities and has the potential to be developed as a natural anticancer agent.

수 종의 한약제제가 조혈작용에 미치는 영향 (Effects of Korean Traditional Medicine on Murine Hematopiesis (Regulation of Hematopoietic Cytokine & $CD34^{+}$ cell Expression))

  • 전재현;김영철;이장훈;우흥정
    • 대한한의학회지
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    • 제22권3호
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    • pp.156-168
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    • 2001
  • Objectives : To evaluate the diverse actions of stimulation on the hematopoietic system, 4 formulas (KH I, KH 2, KH 3, KH 4) were studied. Method and Result : RT-PCR was performed to measure the gene expression of hematopoietic cytokines (TPO, GM-CSF, SCF, IL-3). When bone marrow cells were treated with KH 1, 2, 3, 4, the gene expressions of TPO, SCF, IL-3, and GM-CSF were increased. Flow cytometric analysis was performed to measure the expression of CD34+ cell activity. After 72 hrs culture supplemented with KH 1, 2, 3, 4, the percent of CD34+ cell of KH 2, 3, 4 were increased. To measure the expression of colony forming units - granulocyte erythrocytes, macrophages, megakaryocytes (CFU-GEMM) and burst forming unit-erythroid (BFU-E), semisolid clonogenic assay was performed. After 14 days of culture the number of CFU-GEMM and BFU-E of KH I, 2, 3, 4 were significantly increased compared to those of EPO groups (KH 1 P<0.0l, KH 2 P<0.05, KH 3 P<0.001, KH 4 P<0.0l). To determine the intracelluar TPO expression by KH 3, KH 4 in bone marrow cells, intracelluar staining and flow cytometric analysis were performed. After 24 hrs cultures, the TPO expression of the KH 3 and KH 4 treated groups were increased over those of the controlled groups (control : 50%, KH 3 : 87%, KH 4 : 78%). Conclusion : These results suggest that KH I, KH 2, KH 3, KH 4 have hematopoietic effects through increasing the production of hematopoietic cytokines and stimulating the activity of $CD34^{+}$ cells. This study also shows that KH 3 has a more effective hematopoietic effect than KH 1, 2, 4. These results suggest that the formulas (KH I, 2, 3, 4) can be applied to the patients with inappropriate hematopoietic system, and that KH 3 can be the most effective formula among these 4 in treating bone marrow disease in clinics.

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번행초 추출물의 항염증 및 종양억제 효과 (Anti-inflammatory and Anti-tumor Effects of Tetragonia tetragonoides Extracts)

  • 최혜정;이성태;권기석;주우홍
    • 한국미생물·생명공학회지
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    • 제43권4호
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    • pp.391-395
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    • 2015
  • 본 연구에서는 번행초의 추출물과 분획물의 항염증 효과 및 항암 활성에 대해 연구하였다. 번행초의 항염증 활성은 $1{\mu}g/ml$ LPS로 유도되는 NO 및 IL-$1{\beta}$의 생성이 억제됨을 통해 확인되었다. IL-6 및 $TNF-{\alpha}$$1{\mu}g/ml$ LPS로 자극된 RAW264.7 세포에서 다당류 추출물을 처리한 후 생성이 감소되었다. 나아가 HX 분획물은 GM-CSF의 생성을 강력하게 억제하는 것으로 확인되었다. 또한 Sarcoma 180을 미리 주사한 ICR 마우스에 MeOH과 다당 추출물을 100 mg/kg/day 복강내 투여시 16.67%의 수명 연장 효과가 관찰되었다. 이상의 결과는 번행초를 이용하여 효과적인 항염증 및 항암제 개발을 위한 사전조사로써 중요한 결과를 제공한다.

Effects of Culture Dimensions on Maintenance of Porcine Inner Cell Mass-Derived Cell Self-Renewal

  • Baek, Song;Han, Na Rae;Yun, Jung Im;Hwang, Jae Yeon;Kim, Minseok;Park, Choon Keun;Lee, Eunsong;Lee, Seung Tae
    • Molecules and Cells
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    • 제40권2호
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    • pp.117-122
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    • 2017
  • Despite the fact that porcine embryonic stem cells (ESCs) are a practical study tool, in vitro long-term maintenance of these cells is difficult in a two-dimensional (2D) microenvironment using cellular niche or extracellular matrix proteins. However, a three-dimensional (3D) microenvironment, similar to that enclosing the inner cell mass of the blastocyst, may improve in vitro maintenance of self-renewal. Accordingly, as a first step toward constructing a 3D microenvironment optimized to maintain porcine ESC self-renewal, we investigated different culture dimensions for porcine ICM-derived cells to enhance the maintenance of self-renewal. Porcine ICM-derived cells were cultured in agarose-based 3D hydrogel with self-renewal-friendly mechanics and in 2D culture plates with or without feeder cells. Subsequently, the effects of the 3D microenvironment on maintenance of self-renewal were identified by analyzing colony formation and morphology, alkaline phosphatase (AP) activity, and transcriptional and translational regulation of self-renewal-related genes. The 3D microenvironment using a 1.5% (w/v) agarose-based 3D hydrogel resulted in significantly more colonies with stereoscopic morphology, significantly improved AP activity, and increased protein expression of self-renewal-related genes compared to those in the 2D microenvironment. These results demonstrate that self-renewal of porcine ICM-derived cells can be maintained more effectively in a 3D microenvironment than in a 2D microenvironment. These results will help develop novel culture systems for ICM-derived cells derived from diverse species, which will contribute to stimulating basic and applicable studies related to ESCs.