• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2003년도 생물공학의 동향(XII)
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• pp.105-108
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• 2003

To enhance the performance of a serum-free medium (SFM) for human thrombopoietin (hTPO) production in suspension cultures of recombinant Chinese hamster ovary (rCHO) cells, several low-cost hydrolysates such as yeast hydrolysate (YH), soy hydrolysate, wheat gluten hydrolysate and rice hydrolysate were tested as medium additives. Among various hydrolysates tested, the positive effect of YH on hTPO production was most significant. When 5 g/L YH was added to SFM, the maximum hTPO concentration in batch culture was 40.41 ${\mu}g/mL$, which is 11.5 times higher than that in SFM without YH supplementation. This enhanced hTPO production in YH-supplemented SFM was obtained by the combined effect of enhanced $q_{hTPO}$ and increased culture longevity. In addition YH supplementation did not affect in vivo biological activity of hTPO. Taken together, the results obtained demonstrate the potential of YH as a medium additive for hTPO production in serum-free suspension cultures of rCHO cells.

• Journal of Plant Biotechnology
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• 제4권1호
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• pp.23-27
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• 2002

To establish an in vitro mass production system of grape anthocyanin pigments through callus and cell suspension culture, the effects of nitrogen source and sucrose on fresh weight and anthocyanin production in cell suspension culture of 'Sheridan' grape level were studied. When the medium was devoid of $NO_3^-$, cell fresh weight was either remained stable (1% sucrose) or slightly decreased with culture time (2,3, and 4% sucrose). When $NH_4^-$ was lacking, 3% sucrose was most favorable for cell growth. When $NH_4^-$ was supplied as N source, the anthocyanin content of 2% sucrose containing medium was maintained 2 times higher than other levels till day 8 in culture, then that of 3 and 4% sucrose which peaked at day 12 thereafter. The anthocyanin content was low than $NO_3^-$-free media. Total anthocyanin content in $NH_4^-$-free medium was just about a half of that of $NH_4^+$ medium. Anthocyanin production of 2% sucrose in $NH_4^+$ medium was maintained about 3-fold till day 8, then decreased thereafter. In $NH_4^+$ medium, pH decreased gradually with final pH of 3.5 to 4.0, while pH in $NH_4^+$-free medium increased with final pH of 6.5 to 7.5.

• Journal of Plant Biotechnology
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• 제3권2호
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• pp.101-106
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• 2001

In order to establish efficient plant regeneration from embryogenic suspension cultures of soybean, Glycine max L, we examined the effects of auxin type and concentration, cytokinin type and concentration, and amino acid type and concentration on the growth of embryogenic clumps from induced callus, and the effect of desiccation of mature somatic embryos obtained from these clumps on the frequency of somatic embryo germination. Embryogenic callus was induced from the edge of the cotyledons cultured on MS medium containing 6% sucrose, 40 mg/L 2,4-D, 0.2% gelrite and pH 5.7. The growth of embryogenic clumps was best in early staged, embryogenic callus that was placed in suspension culture of MS medium containing 5 mg/L 2,4-D and 0.5 mg/L asparagine. Single somatic embryos were isolated from the clumps and plated on the same medium for maturation. When the mature single somatic embryos were desiccated for 96 h, somatic embryo germination came up to approximately 90%. The plantlets germinated after embryos desiccation for 2 weeks were transfered to MS medium containing 3% sucrose,0.2% gelrite and pH 5.7.

• Journal of Plant Biotechnology
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• 제5권2호
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• pp.131-135
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• 2003

Callus and cell suspension cultures of Eurycoma longifolia Jack were initiated from leaves of different trees. The leaf explant of tree Eu9 produced the most calli and also induced high cell biomass in the cell suspension culture. Optimum production of cell biomass could be initiated in proliferating culture medium with a pH of 5.75 prior to autoclaving. The effects of macronutrient inorganic salts of Murashige and Skoog (MS) liquid medium supplemented with X on production of cell biomass of Eurycoma longifolia were also investigated. The highest cell biomass was produced in MS medium containing macronutrients of $21\;mM\;NH_4NO_3,\;12.25\;mM\;KNO_3,\;3.00\;mM\;CaCl_2.2H_2O,\;0.575\;mM\;MgSO_4.7H_2O$, and $1.83\;mM\;KH_2PO_4$. A new medium labeled as TAM was formulated for the production of Eurycoma longifolia cell biomass in the cell suspension culture.

• 한국약용작물학회지
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• 제3권2호
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• pp.96-99
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• 1995

본 실험은 쪽 세포의 현탁배양에서 indirubin 생산을 위한 indole의 처리 시기와 처리 기간 및 첨가량을 찾기 위하여 수행하였으며 얻어진 결과는 다음과 같다. 1. Indole 첨가시 세포 생장량은 억제되었다. 2. Tryptophan 첨가시 세포 생장량의 증가는 크지 않았다. 3. Indole과 L-tryptophan을 고체레지에 첨가한 경우 indole을 첨가한 배지에서 만 indirubin이 검출되었다. 4. Indirubin 생산에 적절한 indole 첨가 농도는 고체배지에서 200mg으로 나타났다. 5. 현탁배양 중 indole 첨가시기를 달리한 결과 배양 20일 후 indole을 첨가한 배지에서 세포와 배지내 indirubin 농도가 높았다. 6. 고체배양보다 현탁배양에서 세포내에 축적된indirubin이 많았으며 배지 내 로 상당량의 indirubin이 유출되었다.

• 한국자원식물학회지
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• 제24권3호
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• pp.280-285
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• 2011

In this study, we established a novel somatic embryogenesis and plant regeneration system through cell suspension culture of white dandelion (Taraxacum coreanum NAKAI.). Embryogenic calli could be initiated from leaf and root explants of sterile seedlings on solid Murashige and Skoog (MS) medium supplemented with 1.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) after 3-week cultures. To proliferate embryogenic calli rapidly, cell suspension culture was performed with transferred to liquid MS medium with various combinations of plant growth regulators (PGRs) including 2,4-D, ${\alpha}$-naphthalene acetic acid (NAA), indole-3-acetic acid (IAA), $N^6$-benzylamino purine (BAP), thidiazuron (TDZ), and kinetin. During suspension cultures, embryogenic calli not only greatly proliferated, but shoot organogenesis also simultaneously occurred from the surface of somatic embryos. Among them, TDZ at lower concentration, 0.1 mg/L produced the highest efficiency of somatic embryo formation and shoot organogenesis. Rooting of embryogenic calli with adventitious shoots was done on solid MS medium containing 0.1 mg/L NAA and 0.3% activated carbon. Nearly 80% of embryogenic calli with shoot organogenesis could be rooted normal. Well-rooted plantlets were transferred into pots under a greenhouse condition, and plants derived from this system appeared phenotypically normal.

• Biotechnology and Bioprocess Engineering:BBE
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• 제10권6호
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• pp.564-570
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• 2005

In the present study, the protective effects of Bcl-2 over-expression in a suspension culture (without any adaptation) and spent medium (low nutrient and high toxic metabolite conditions) were investigated. In the suspension culture without prior adaptation, the viability of the control cell line fall to 0% by day 7, whereas the Bcl-2 cell line had a viability of 65%. The difference in the viability and viable cell density between the Bcl-2 and control cell lines was more apparent in the suspension culture than the static culture, and became even more apparent on day 6. Fluorescence microscopic counting revealed that the major mechanism of cell death in the control cell line in both the static and suspension cultures was apoptosis. For the Bcl-2 cell lines, necrosis was the major mode of cell death in the static culture, but apoptosis became equally important in the suspension culture. When the NS0 6A1 cell line was cultured in spent medium taken from a 14 day batch culture, the control cell line almost completely lost its viability by day 5, whereas, the Bcl-2 still had a viability of 73%. The viable cell density and viability of the Bcl-2 cell line cultivated in fresh medium were 2.2 and 2.7 fold higher, respectively, than those of the control cultures. However, the viable cell density and viability of the Bcl-2 cultivated in the spent medium were 8.7 and 7.8 fold higher, respectively, than those of the control cultures. Most of the dead cells in the control cell line were apoptotic; whereas, the major cell death mechanisms in the Bcl-2 cell line were necrotic.

• 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.

• Archives of Pharmacal Research
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• 제16권3호
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• pp.244-250
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• 1993

Calli and suspension cultures were obtained following inoculation of the explant from leaves of Ginkgo biloba L on the supplemented MS basal medium. The obtained calli and suspension cultured cells were able to produce detectable amounts of ginkgolides which are known as natural specific PAF antagonists. The production of ginkgolides in the calli and suspension cultured celles were identified using GC/MS, GC and HPLC with authentic ocmpounds. Since the production of ginkgolides A and B the calli and suspension cultured cells had been confirmed, effects of types and concentration of plant growth regulators, media and illumination on the induction and growth of the callus were studied. The concentrations of growth regulators for optimal callus were studied. The concentrations of growth regulators for optimal callus induction were studied. The concentrations of growth regulators for optimal callus induction were 1.0 to 2.0 mg/L for NAA and o.1 mg/L for kinetin. The growth of the Callus seemed to be more simnultaed with the combination of NAA and kinetin than NAA and BA with illumination at all concentration ranges of 1.0 to 4.0 mg/l for NAA and o.1 to 1.0 mg/L for kinetin or BA studied. Amogn 8 different media used, the induction rate of callus on Anderson, Eriksson, and Shenk and Hildebrant at 4 weeks after the innoculation was almost the same as that of MS. However, callus was rarely induced on Heller or White medium. Suspension cultures were easily initiated with 3 g of callus (fresh weight) derived from ginkgo leaves on supplemented MS medium. A typical growth curve of suspension cultured cells could be obtained by measuring the fresh weight of the suspension cultured cells at every 3 days. To improve the growth of suspension cultured cells of ginkgo, effects of concentrations of NAA, sucrose, phosphate ions and molar ratio of $NH_{4}^+\;to\;NO_{3}^-$ ions in the culture medium were studied. The maximum growth of the cells was achieved when the culture medium contained 1.0 mg/L of NAA, 30 g/L sucrose, 1.75 mM phosphate ions and 1:5 molar ratio of $NH_{4}\;to\;NO_{3}^-$ ions.

• Journal of Plant Biotechnology
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• 제6권2호
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• pp.125-130
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• 2004

Callus and cell suspension cultures of Eurycoma longifolia Jack could be an alternative supply of 9-hydroxycanthin-6-one and 9-methoxycanthin-6-one. The callus tissues were initiated from leaves of different trees. The friable calli were used for the preparation of the cell suspension cultures of E. longifolia. The leaf explant of tree Eu-9 produced the most callus and also induced high cell biomass in the cell suspension culture, but it produced low quantity of 9-methoxycanthin- 6-one and 9-hydroxycanthin-6-one. The leaf explant from tree Eu-8 produced low quantity of callus and cell biomass, but produced the highest quantity of 9-methoxycanthin- 6-one and 9-hydroxycanthin-6-one. Optimum production of cell biomass was obtained on cell culture medium with pH 5.75 prior to autoclaving, but high alkaloids content could be induced in culture medium in acidic condition with pH 4.75 and 5.25 prior to autoclaving.