• 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.

• 한국자동차공학회논문집
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• 제6권3호
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• pp.154-160
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• 1998

Ride quality of medium truck became a very important factor in the suspension design, to the demand of more comfortable ride of passengers. This study describes how to determine and evaluate design parameters related to the chassis suspension system with time and frequency analysis. The spring stiffness and damping force of the chassis suspension system were obtained by observing the vertical acceleration PSD. The simulation was carried out on various road profiles, which was suggested by ISO. The pitching motion of the medium size truck was observed to improve the ride quality. A computer simulated truck model was constructed using DADS, a commercial dynamic analysis software, in order to simulate the truck motions. From the analyzed process of suspension parameters, it was concluded that the spring and the shock absorbers affect the pitching of the vehicle. In order to validate the computer simulated truck model, a physical prototype was constructed and tested.

• 한국정밀공학회:학술대회논문집
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• 한국정밀공학회 2010년도 춘계학술대회 논문집
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• pp.1451-1452
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• 2010

• Journal of Plant Biotechnology
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• 제37권2호
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• pp.228-235
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• 2010

The establishment of cell suspension culture and plant regeneration of the halophytic Leymus chinensis (Trin.) are described in this study for the first time. Callus induction solid medium containing Murashige and Shoog (MS) basic salt, $2.0\;mg\;l^{-1}$ 2,4-dichlorophenoxyacetic acid (2,4-D), and $5.0\;mg\;l^{-1}$ L-glutamic acid with $30.0\;g\;l^{-1}$ sucrose and $4.0\;g\;l^{-1}$ gelrite for solidification induced the highest rate of cell division in Type 1 callus among calli of various types. Liquid medium with the same hormone distribution was therefore, used for cell suspension culture from Type 1 callus. Over a 30 d suspension culture at 100 rpm, great amounts of biomass were accumulated, with 71.07% average daily increment and 22.32-fold total fresh weight increment. Comparison of before and after suspension culture, the distribution of different size callus pieces and the maintenance of callus type were basically unaltered, but a slight increase in relative water contents was observed. To induce the potential of plant regeneration, the directly transferring on plant regeneration solid medium containing MS basic salt, $0.2\;mg\;l^{-1}$ $\alpha$-naphthalene acetic acid (NAA), $2.0\;mg\;l^{-1}$ kinetin (Kn), and $2.0\;g\;l^{-1}$ casamino acid and indirectly transferring were simultaneously performed. Even now growth rates of suspension-derived callus on solid medium were approximately half of those of Type 1 callus, but faster somatic embryogenesis was observed. Rooting of all regenerated shoots was successfully performed on half-strength MS medium. All plants appeared phenotypically normal.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2003년도 생물공학의 동향(XIII)
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• pp.171-173
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• 2003

본 연구에서는 생분해성 고분자 나노입자를 이용하여 부착성 세포의 세포군집 (cell aggregates) 형성을 촉진시켜 무혈청 배지에서 3차원적 부유 배양하는 방법을 개발했다. 생분해성 고분자 나노입자의 사용은 무혈청 배지 부유배양에서 부착성 동물세포인 HEK 293 세포의 세포군집 형성과 세포증식(나노입자를 사용하지 않은 대조군과 비교하여 2배 이상)을 촉진하였다. 일반적으로 무혈청배지 부유배양에 세포를 적응(adaptation)시키는 데에는 시간이 오래 걸리고 많은 비용이 드는데, 이 연구에서 개발된 방법은 이러한 세포적응 공정이 필요없다. 이 배양법은 여러 부착성 동물세포의 산업적 대량배양에 유용하게 사용될 수 있을 것이다.

• 한국약용작물학회지
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• 제12권5호
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• pp.366-371
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• 2004

Aconitine alkaloids produced from cell suspension cultures of Aconitum napellus for the first time. The effects of various culture conditions on cell biomass and aconitine accumulation in cell suspension cultures were investigated. Suspension cell cultures of A. napellus were established by transferring callus tissues from leaf explants onto liquid MS medium supplemented with $1\;mg/l$ NAA and $0.1\;mg/l$ kinetin. Among the culture media tested, MS medium had a pronounced effect on cell growth and aconitine accumulation. The maximum dry cell weight was obtained at inoculum size of 3 g (FCW) per flask and in MS medium supplemented with 5% sucrose after 8 weeks. The addition of salicylic acid (SA) and yeast extract (YE) in the MS medium enhanced aconitine accumulation. Using a proper combination of culture condition and supplements, aconitine content could reach 0.043% (dry weight basis), that was $2.5{\sim}3$ fold higher that detected in control cultures.

• Biotechnology and Bioprocess Engineering:BBE
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• 제11권2호
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• pp.154-159
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• 2006

Interleukin-18 (IL-18), otherwise known as interferon-gamma-inducing factor (IGIF), is one of several well characterized and important cytokines that contribute to host defenses. The complementary DNA (cDNA) of mature human interleukin-18 gene (hIL-18) was fused with the signal peptide of the rice amylase 1A gene (Ramy1A) and introduced into the plant expression vector under the control of a duplicated CaMV 35S promoter. The recombinant plasmid was transformed into tobacco (Nicotiana tabacum L. cv Havana) using the Agrobacterium-mediated transformation method. The integration of the hlL-18 gene into the genome of transgenic tobacco plants was confirmed by polymerase chain reaction (PCR) amplification and its expression was observed in the suspension cells that were derived from the transgenic plant callus by using Northern blot analysis. The hlL-18 protein was detected in the extracts of the transgenic callus and in the medium of the transgenic tobacco suspension culture by using immunoblot analysis. Based upon enzyme-linked immunosorbant assay (ELISA) results, the expression level of the hlL-18 protein approximated $166{\mu}g/L$ in the suspension culture medium. Bioassay results from the induction of $interferon-{\gamma}$ from a KG-1 cell line indicated that the hlL-18 secreted into the suspension culture medium was bioactive.

• Plant Biotechnology Reports
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• 제2권1호
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• pp.33-39
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• 2008

In this paper, we would like to show unexpected morphogenic potential of cell suspensions derived from seedling explants of Gentiana kurroo (Royle). Suspension cultures were established with the use of embryogenic callus derived from seedling explants (root, hypocotyl and cotyledons). Proembryogenic mass proliferated in liquid MS medium supplemented with $0.5mg\;l^{-1}$ 2,4-D and $1.0mg\;l^{-1}$ Kin. The highest growth coefficient was achieved for root derived cell suspensions. The microscopic analysis showed differences in aggregate structure depending on their size. To assess the embryogenic capability of the particular culture, 100 mg of cell aggregates was implanted on MS agar medium supplemented with Kin ($0.0-2.0mg\;l^{-1}$), $GA_3$ ($0.0-2.0mg\;l^{-1}$) and AS ($80.0mg\;l^{-1}$). The highest number of somatic embryos was obtained for cotyledon-derived cell suspension on $GA_3$-free medium, but the best morphological quality of embryos was observed in the presence of $0.5-1.0mg\;l^{-1}$ Kin, $0.5mg\;l^{-1}$ $GA_3$ and $80.0mg\;l^{-1}$ AS. The morphogenic competence of cultures also depended on the size of the aggregate fraction and was lower when size of aggregates decreased. Flow cytometry analysis reveled luck of uniformity of regenerants derived from hypocotyl suspension and 100% of uniformity for cotyledon suspension.

• 한국약용작물학회지
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• 제12권6호
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• pp.442-447
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• 2004

Cell growth and gomisin J production by suspension cultures of Schisandra chinensis Baillon were investigated under various culture media, initial sucrose concentrations, shaking speeds, and inoculum sizes. Callus was induced from in vitro cultivated leaf segments on MS medium supplemented with $1\;mg/{\ell}$ NAA. The maximum dry cell weight of 2.23 g was obtained at inoculum size of 0.5 g fresh cell weight and in MB5 medium supplemented with $1\;mg/{\ell}$ NAA, 3% sucrose after 8 weeks. The production of gomisin J in suspension cell cultures was maximized in WPM medium containing 5% sucrose. The shaking speed for maintaining maximal cell dry weight was 100 rpm while the best shaking speed for gomisin J accumulation was 140 rpm.

• 식물조직배양학회지
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• 제26권4호
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• pp.289-291
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• 1999

발아 7일된 오이 (Cucumis sativus L.) F$_1$ 5품종과 순계 4품종의 하배축 절편을 1mg/L 2,4-D가 첨가된 MS고체배지에 치상하여 3주간 배양하였다. F$_1$ 1품종과 순계 2품종에서 발달한 캘러스의 8%이하에서 체세포배가 발생하였다. 체세포 배 절편을 동일한 배지에 4주간 배양하여 배발생캘러스를 유도하였다. 배발생캘러스는 동일조성의 MS액체배지에 현탁되어 증식하였다. 액체배지에서 배발생캘러스는 수많은 체세포 배로 발달하였으며, 생장조절제가 첨가되지 않은 MS기본배지에서 95%이상의 체세포배가 식물체로 발아하였다. 재분화된 식물체는 화분에서 생육되어 개화 후 착과하였다.