• The Korean Journal of Ecology
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• 제20권3호
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• pp.169-173
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• 1997

The measurement of cell death constant in Anabaena flos-aquae was tested by the Live/Dead BacLight Viability kit(Molecular Probes Co., Seatle, WA). When the Live/Dead BacLight Viability kit was applied to Anabaena flos-aquae, the cells with intact cell membranes(live cells) stained fluorescent green, while the cell with damaged membranes(dead cells) stained fluorescent red and the background remained virtually nonfluorescent. The rations of live : dead cells in the cell suspension were controlled artifically and Live/Dead BacLight Viability kit was applied to them. The ratios of green:red fluorescent cells in the cell suspension were the same as those of live : dead cells controlled artifically. It was also approved by the fluorescence emission. The cell death constant was measured in the P-limited Anabaena flos-aquae chemostal culture in the N-fixing and $KNO_3-supplied$ conditions. The culture in N-fixing chemostat had a dead cell proportion of 1.2% at the growth rate of 0.7/day and increased to 2.6% at the growth rate of 0.3/day. The cell death constant of N-fixing culture was 0.008/day.There was a same trend in the $KNO_3-supplied$ chemostat culture. The proportion of dead cell was 1.6% of dead cell proportion at the growth rate of 0.7/day and increased to 4.3% at the growth rate of 0.3/day.

• Journal of Plant Biology
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• 제33권3호
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• pp.189-196
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• 1990

Suspension cell lines have been newly established from the calli derived from the immuature embryo culture of hexapolid (Triticum aestivum var. sicco), tetrapolid (T. durum) and diploid (T. tauchii or Aegilops squarrosa) wheat species. The chromosomal variation in suspension cultured cell lines was examined and old cell line, C82d, established from T. aestivum var. copain was also used. New method using 1-bromonaphthalene for metaphase rapping of suspension cells was developed. Variation in chromosome number was observed among all the suspension lines. Cells with doubled chromosome number and deleted chromosome were also observed. Extensive structural changes in chromosome were found in C82d line. Chromosome aberrations showed loss of chromosome arms and chromosome segment. The mean chromosome number in suspension cells of T. aestivum var. sicco was 40, in C82d line 33, in T. durum 28 and in T. tauchii 14. The stability of chromosome in suspension cells of diploid and tetrapolid wheats was higher than that of hexaploid wheat.

• 식물조직배양학회지
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• 제21권1호
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• pp.47-53
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• 1994

본 실험은 포도와 미국자리공의 세포현탁배양계에서 세포생장과 athocyaninn 및 betacyanin 색소생성에 미치는 광의 영향을 조사하였다. 그 결과, 포도의 세포현탁배양계에서의 세포증식 패턴은 광의 효과가 미약하였으나, 미국자리공의 경우는 암배양에 비하여 광조사구에서 뚜렷한 세포증식 패턴을 나타내었다. 포도세포는 12일간의 배양기간중 한번의 생장 피크를 보여주는데 비하여 미국자리공의 배양세포는 두 번의 증식 피크를 보여 광이 세포의 생장 및 분열 주기를 촉진시킬 뿐만 아니라 색소의 축적도 촉진시키는 것으로 나타났으며, 미국자리공 배양세포는 배양 후 4일째에 액포화에 따른 색소의 축적이 확인되었다. 한편, 색소생성에 미치는 광의 영향은 포도 현탁배양계에서는 배양후 6일째부터 생성되기 시작하여 배양 후 10일 전후에 최고치를 보이는데 비하여, 미국자리공의 현탁배양계에서는 세포증식 패턴에 따라 배양 후 4일째와 8일째에 두 번의 피크를 보였다. 그러나 이들 두 가지 모두 암배양하에서는 거의 색소 생합성이 이루어지지 않고 광상태하에서만 색소의 축적을 볼 수 있었다. 한편, 포도 세포배양계에서는 배양 후 4일째에 처리한 광은 약간의 효과가 있었으나, 배양 후 7일째 12시간 이상의 광에 의하여 색소생성이 크게 촉진되는 것을 알 수 있었다. 미국자리공의 경우에는 배양 후 7일 째보다는 오히려 배양 후 4일째에 광이 더욱 효과적이었다. 이상의 사실은 이들 두 가지 색소의 생합성과정에는 광이 절대적으로 필수적이지만 광을 필요로 하는 시기 및 요구하는 광량 등이 서로 다르다는 것을 시사해 준다.

• 한국생명과학회:학술대회논문집
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• 한국생명과학회 2002년도 제38회 학술심포지움
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• pp.48-60
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• 2002

Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a cytokine that stimulates the production of granulocytes, macrophages, and white blood cells. The effects of osmotic pressure on secretion of human GM-CSF into the culture medium were investigated in suspension cultures of transgenic tobacco cells. An increase in osmotic pressure caused by the addition of mannitol decreased the cell size index, with the effect being more pronounced when cells were measured wet rather than dry. Increased osmotic pressure enhanced the secretion of hGM-CSF. At 90 g/L mannitol, the maximum concentration tested, hGM-CSF was present in the culture medium at 980 ug/L. As the concentration of mannitol increased, the total amount of protein secreted also increased, but was disproportionately enriched in GM-CSF NaCl, another osmoticum, had very similar effects on cell growth and hGM-CSF production, but did not cause enrichment for hGM-CSF Additionally, protein-stabilizing polymer was added to culture broth to enhance stability of secreted recombinant protein. Finally, above two method were applied together to maximize the productivity.

• KSBB Journal
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• 제19권5호
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• pp.374-380
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• 2004

Production of therapeutic proteins by transgenic plant cell suspension cultures is an attractive system alternative to the other expression system. However, plant cell cultures have shown low expression level of foreign proteins and decreased cell viability by the changes of culture conditions. Therefore, it is necessary to enhance cell viability during the culture period. In this study, a quantitative analysis technique was designed to measure relative cell viability for plant suspension cells which have cell wall and aggregates. It was found that the programmed cell death of plant cells by apoptosis was essentially linked with the apoptotic pathway of animal cells. Therefore, effects of nicotinamide, 3-aminobenzamide and antioxidants on cell viability and apoptosis were examined in transgenic Nicotiana tabacum cells producing hGM-CSF. With those additives, cell viability could be maintained and apoptosis could be redued. In the result, the extracellular production of hGM-CSF could be enhanced 2.5 fold. It was also found that the supplementation of glutathione and ascorbic acid suppressed both the cold stress-induced decrease in cell viability and the increase of total genomic DNA fragmentation.

• Biotechnology and Bioprocess Engineering:BBE
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• 제8권2호
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• pp.135-141
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• 2003

The human granulocyte-macrophage colony stimulating factor (hGM-CSF) gene was introduced into tobacco plants. The cell suspension culture was established from leaf-derived calli of the transgenic tobacco plants in order to express and secrete a biologically active hGM -CSF. The recombinant hGM-CSF from the transgenic plant cell culture (prhGM-CSF) was identified as a yield of about 180 ${\mu}$g/L in the culture filtrate, as determined by ELISA. The addition of 0.5 g/L polyvinylpyrrolidone (PVP) to the plant cell culture medium both stabilized the secreted prhGM-CSF and increased the level of production approximately 1.5-fold to 270 ${\mu}$g/L. The biological activity of the prhGM-CSF was confirmed by measuring the proliferation of the hGM-CSF-dependent cell line, TF-1. Interestingly, the specific activity of the prhGM-CSF was estimated to be approximately 2.7 times higher than that of a commercially available preparation from E. coli.

• 한국환경과학회지
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• 제15권12호
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• pp.1193-1197
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• 2006

This study was to examine the variations of chromosome number and the ranges of variety in the suspension culture of ginseng (Panax ginseng C. A. Meyer) callus cell, and the effect of plant hormones for the chromosome aberration. Plant hormones added with MS medium in the suspension culture were 2,4-D, kinetin, and 2,4-D+kinetin and concentration of the plant hormones were $1000{\mu}M$ and $0.1\;{\mu}M$ respectively. As a result of these experiment the following conclusion has been obtained. Media contained with 2,4-D+kinetin in $10{\mu}M$ concentration was very effective in the suspension culture result from 26.4% mitosis frequency, and found the various variation of chromosome number. Variety of chromosome number was diversed ($9\sim110$), espicially frequency of hypohaploid and hyperhaploid cells were very higher than hyperdiploid cells. In this experiments, it is suggested that $10{\mu}M$ 2,4-D+kinetin added with medium in the suspension culture of ginseng callus was effect in the variations of chromosome number.

• 대한의생명과학회지
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• 제19권1호
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• pp.41-47
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• 2013

Pluripotent stem cells (PSCs) have lots of potential in biomedical sciences owing to its potential to differentiate into any kind of cells in the body. However, it is still a challenge to culture PSCs on a large scale for application to regenerative medicine. Herein, we introduce a synthetic polymer that enables large-scale suspension culture of human PSCs. By employing suspension culture, it became unnecessary to use conventional substrata such as mouse embryonic fibroblast (MEF) or Matrigel$^{TM}$, which are believed to be main causative sources of xenogeneic contamination in cultured human PSCs in vitro. Human PSCs were cultured in the medium in which elastin-like polypeptide (ELP) dissolved. The ELP in the medium became harden as temperature increases by transforming the medium into a semi-solid gel that supported growth of human PSCs in suspension. Gel-sol transition temperature of ELP can be adjusted by modifying the peptide sequence in which 5 amino acids, Val-Pro-Gly-Xaa-Gly, repeated sequentially. We constructed 3D suspension media having transition temperature around $33{\sim}35^{\circ}C$ using an ELP consisted of 40, 60, or 80 repeats of a monomer, which was Val-Pro-Gly-Val-Gly. Among the ELPs, ELP80 was chosen as the best ELP to support growth of human PSCs in suspension culture. This result suggests that the ELP80 can be a medium component for culturing human PSCs in large-scale.

• 아시안잔디학회지
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• 제23권2호
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• pp.345-352
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• 2009

한국들잔디는 운동경기장이나 골프 코스 등에 폭넓게 쓰이는 난지형 잔디이다. 많은 품종은 영양체로 번식한다. 이 연구는 배아세포 유도와 식물체 재생을 위한 적정 배양액 개발과 배양조건을 연구하여, 한국잔디의 육종과 번식을 위한 세포배양 체계 확보를 목적으로 하였다. 그 결과 $Cu^{++}\;2.5mgL^{-1}$를 유도 배양액에 첨가하는 것이 캘러스 유도를 위한 최적 농도 조건 이였음을 보여주었다. Sub-culture의 횟수의 증가는 캘러스의 질을 향상시켰다. 세포 부유배양을 위한 최적 용량은 2.5에서 10mL 사이였으며 본 연구에서 58%의 식물체 재생산비율을 보였다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2001년도 추계학술발표대회
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• pp.215-216
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• 2001

Mouse monoclonal antibody(mAb) with an antigen specificity for major histocompatibility complex class Il(MHC class II) was produced and secreted from tobacco cell suspension culture by successive sexual crossesu. Expression and secretion of assembled antibody was observed in transgenic tobacco cell suspension culture by wetern blot analysis.