• Journal of Plant Biotechnology
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• 제3권2호
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• pp.83-88
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• 2001

A cell culture system of poplar (Populus alba x P.glandulosa) was established to test four different methods for evaluation of cellular stresses. Two different kinds of stresses were given to the cultures by adding either Pb(NO$_3$)$_2$ or paraquat and the cellular responses were monitored during a week period. While fresh weight reduction was observable in two days after the treatment of Pb(NO$_3$)$_2$, such changes were apparent only in later stage in paraquat treated cultures. Cells in paraquat treated cultures in the first 3 days showed no alteration in fresh weight as compared to untreated cultures, but had their MTT reducing activities completely inhibited. Neither Evans blue staining nor ion conductivity of the medium was consistent with fresh weight changes of the cultures. Overall, cell clumps formed during suspension culture appeared to interfere with staining and washing reactions and thus cause the assays unreliable. Among the four methods examined, fresh weight changes and MTT reducing activity appeared to be the most reliable and consistent.

• Journal of Plant Biology
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• 제36권3호
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• pp.211-218
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• 1993

Plant regeneration was accomplished from protoplast culture of rice (Oryza sativa L. cv. Taebaeg). Embryogenic callus was induced from mature seed on MS medium containing 5 mM proline, 2.5 mg/L 2,4-D, 30 g/L sucrose in the dark at 28$^{\circ}C$ and used to establish embryogenic cell suspension culture. Suspension cells were subcultured every one week in N6 medium supplemented with 5 mM proline, 200 mg/L casein hydrolysate, 2.5 mg/L 2,4-D and amino acids of AA medium. Suspension cultures were composed of cells that were densely cytoplasmic, potentially embryogenic and were at least maintained for more than 6 months in liquid medium. Protoplasts were isolated from fast-growing suspension culture cells and cultured in a slightly modified KpR medium by mixed nurse culture. Isolated protoplasts began to divide within 5~7 days and thereafter, protoplast-derived calli were sequentially transferred to callus proliferating medium that soft agar MS medium contained 2 mg/L 2,4-D and produced distinct embryogenic cells. Microcolonies were then transferred to solid medium which consisted of MS medium containing 5 mg/L kinetin, 1 mg/L NAA, 1 mg/L ABA, 30 g/L sucrose and 10 g/L sorbitol under fluorescent light. Mulitple shoots of 4~5 per callus emerged and were transferred to hormone-free MS medium for root initiation. Thereafter, The plantlets were transferred to pots of soil to mature in the culture room.

• Archives of Pharmacal Research
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• 제17권1호
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• pp.48-50
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• 1994

Three p-coumaroylamino acids (p-CAAs) were isolated from the yeast-elicited Ephedra distachya cultures by consecutive purification using XAD_2, silicagel and RP-HPLC. Retention times on HPLC as well as their UV, IR, NMR and MS spectral data indicated that the yeast-induced p-CAAs wre p-coumaroyl--D-valine, p-coumaroyl-D-serine and p-coumarouyl-D-threonine, respectively. The structures of p-CAAs were confirmed by the comparison of their physico-chemical properties 3with those of synthetic ones. They were isolated and identified for the first time from natural products and supposed to be accumulated as phytoalexins of Ephedra.

• 한국식물생명공학회:학술대회논문집
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• 한국식물생명공학회 2005년도 추계학술대회 및 한일 식물생명공학 심포지엄
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• pp.112-119
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• 2005

Efficient plant regenerationsystem from cell suspension cultures was established in D. acicularis (2n = 90) by monitoring ploidy level and visual selection of the cultures. The highly regenerable cell lines selected maintained original ploidy level and consisted of compact cell clumps with yellowish color and relatively moderate growth, suggesting that it is possible to select visually the highly regenerable cell lines with the original ploidy level. All the regenerated plantlets from the highly regenerable cell cultures exhibited normal phenotypes and no variations in ploidy level were observed by flow cytometry (FCM) analysis.

• 생명과학회지
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• 제9권4호
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• pp.408-413
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• 1999

Extracellular capsidiol, sesquiterpenoid phytoalexin, in the medium of pepper (Capsicum annuum L.) suspension cells was not identified from control cells, but highly accumulated in the elicitor-induced cells within 6 hours after the addition of 0.05$\mu\textrm{g}$/$m\ell$ cellulase. Capsidiol production in elicitor-induced cells was markedly suppressed by cytochrome P450 inhibitors, such as ancymidol and ketoconazole demonstrating that biosynthesis of capsidiol is catalyzed by at least on hydroxylation enzyme in the biochemical pathway. Based on protein electrophoresis, two bands, 23.0kDa and 27.5kDa, were identified as newly synthesized polypeptides in the elicitor-induced suspension cells, suggesting that pepper cells which were subjected to elicitor treatment activate specific gene(s) for capsidiol biosynthesis in cultured cells.

• 한국동물학회지
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• 제19권3호
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• pp.113-122
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• 1976

非同時化시킨 HeLa $S_3$ 細胞에 있어 MMS에 의한 $^3 H$-Thymidine Uptake率은 MMS의 頻度증가에 따라 직선적으로 감소하며, BUdR 혹은 IUdR과 二重處理할 경우 그 감소율은 더욱 증가한다. MMS에 의한 $^3 H$-Thymidine Uptake은 thymidine 二重 處理후 얻은 同時化된 HeLa $S_3$ 細胞의 $G_2$ 時期에서 檢出된다. BUdR과 IUdR은 MMS에 의한 $^3 H$-Thymidine Uptake率을 더욱 증가시키며 IUdR은 BUdR에 비해 더효과적인 感受性 物質로 판명되었다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2005년도 생물공학의 동향(XVII)
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• pp.392-392
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• 2005

• 한국원자력학회:학술대회논문집
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• 한국원자력학회 2005년도 춘계학술발표회
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• pp.994-995
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• 2005

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2003년도 생물공학의 동향(XIII)
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• pp.351-355
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• 2003

Screening 실험을 통하여 결정된 주요인자인 sucrose, nitrogen, 배양온도로 인자의 최적수준을 결정하는 표면반응법중 하나인 Box-Behnken design을 수행하였다. 각 인자의 상관관계를 통하여 sucrose는 90 g/L, nitrogen은 41 mM, 온도는 $22^{\circ}C$가 최적수준으로 결정이 되었으며 확인실험을 통하여 대조구보다 세포생장이 증가하였을 뿐만 아니라 hGM-CSF의 생산도 약 2배 정도 증가되었음을 확인하였다. 이것은 고농도의 sucrose로 인해 배지내로의 hGM-CSF의 분비가 촉진되었기 때문이며 또한 저온으로 인해 분비된 hGM-CSF를 분해하는 protease 활성이 감소되었기 때문인 것으로 사료된다.

• Journal of Microbiology and Biotechnology
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• 제14권6호
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• pp.1129-1133
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• 2004

The effects of Pluronic F-68, a non-ionic surfactant, on the growth and physical characteristics of Digitalis lanata suspension cultures were investigated in aqueous two-phase systems (ATPSs) composed of $4.5\%$ polyethylene glycol (PEG) 20,000 and $2.8\%$ crude dextran. In the range of 0.1-10.0 g $1^{-1}$, Pluronic F-68 enhanced the maximum cell density in a medium with ATPSs, even though Pluronic F-68 did not affect cell growth in a normal growth medium. In terms of physical properties of ATPSs with cell suspension cultures, 0.2 g $1^{-1}$ of Pluronic F-68 reduced viscosity by up to $40\%$, while 0.1 g $1^{-1}$ of Pluronic F-68 significantly enhanced the oxygen transfer rate. In addition, we successfully performed aqueous two-phase cultivation in a 5-1 stirred tank bioreactor with 0.5 g $1^{-1}$ of Pluronic F-68, and discovered that cell growth in ATPSs was similar to that in normal growth medium.