• Journal of Biomedical Engineering Research
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• v.36 no.6
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• pp.296-301
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• 2015

Explant culture condition of dorsal root ganglion have been used to investigate the pathophysiology of peripheral nerve injury, while applying for the various clinical symptom such as trauma, pressure, and stretch. However, explant culture is usually contaminated by mitotic cells, which may observed as a newly divided cells including fibroblast or glia. The mitotic cells could be able to interrupt and change the cell signaling that make it difficult to avoid detrimental effects during the experiments. To eliminate mitotic cells, anti-mitotic reagents like mixture of uridine and 5-fluorodeoxyuridine or cytosine arabinoside were added to the cultures on the following day, but there is no research that investigate viability of anti-mitotic reagent in dorsal root ganglion explant culture. In this study, we investigate inhibition effect of cytosine arabinoside to mitotic cells in dorsal root ganglion explant culture. Also we visualized and analyzed anti-mitotic effect and toxicity of cytosine arabinoside in various concentration condition. This dorsal root ganglion explant culture condition can be applied to research that effect and mechanism of various stimulation and chemical application which affect peripheral nerve regeneration.

• Journal of Plant Biotechnology
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• v.42 no.3
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• pp.228-234
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• 2015

Using either the apical or axillary bud of the endangered species Abeliophyllum distichum Nakai, we tested the effect of bud position and culture method on shoot proliferation and rooting. In shoot proliferation, the axillary bud explant was more effective than the apical bud and the effect was fostered by BA treatment, whereas no differences were observed in shoot elongation by the explant position. Spontaneous rooting was observed in the MS basal medium and resulted in conspicuous differences in the explant position : more than 80% in apical bud explant and 28% in axillary bud explant was achieved, respectively. The positional effects were also observed in BA pre-treatments: generally vertical culture method appeared to be better in shoot proliferation, growth, and rooting than that of the horizontal culture method regardless of the BA pre-treatment duration. The highest shoot multiplication was achieved through the vertical culture method with axillary bud explant, whereas the best shoot elongation and rooting was obtained using the vertical culture method with the apical bud explant. Apical bud explant was superior to axillary bud explant in ex vitro micro-cuttings and revealed a significant difference in shoot growth and root development. The above results suggest that explant position and culture method influence the efficiency of micropropagation for a rare and endangered plant Abeliophyllum distichum.

• Korean Journal of Medicinal Crop Science
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• v.20 no.6
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• pp.493-499
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• 2012

We have established adventitious root culture systems of Codonopsis lanceolata, Codonopsis pilosula and Codonopsis ussuriensis. Root segments of C. lanceolata were the best explants for induction of adventitious roots and the number of adventitious root for explant was highest on solid medium with $0.5mg/{\ell}$ NAA and produced $18.8{\pm}1.9$ roots per explant. Root segments of C. pilosula were the best explants for induction of adventitious roots and the number of adventitious root for explant was highest on solid medium with $1.0mg/{\ell}$ NAA and produced $8.5{\pm}1.8$ roots per explant. Leaf segments of C. ussuriensis were the best explants for induction of adventitious roots and the number of adventitious root for explant was highest on solid medium with $0.5mg/{\ell}$ NAA and produced $7.8{\pm}0.4$ roots per explant. In liquid culture, the best production of adventitious root (fresh weight) was obtained in 1/2 MS medium with $1.0mg/{\ell}$ NAA. This study demonstrated for the first time to produce adventitious roots in C. pilosula and C. ussuriensis.

• International Journal of Stem Cells
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• v.17 no.3
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• pp.330-336
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• 2024

Mesenchymal stem cells in the dental tissue indicate a disposition for differentiation into diverse dental lineages and contain enormous potential as the important means for regenerative medicine in dentistry. Among various dental tissues, the dental pulp contains stem cells, progenitor cells and odontoblasts for maintaining dentin homeostasis. The conventional culture of stem cells holds a limit as the living tissue constitutes the three-dimensional (3D) structure. Recent development in the organoid cultures have successfully recapitulated 3D structure and advanced to the assembling of different types. In the current study, the protocol for 3D explant culture of the human dental pulp tissue has been established by adopting the organoid culture. After isolating dental pulp from human tooth, the intact tissue was placed between two layers for Matrigel with addition of the culture medium. The reticular outgrowth of pre-odontoblast layer continued for a month and the random accumulation of dentin was observed near the end. Electron microscopy showed the cellular organization and in situ development of dentin, and immunohistochemistry exhibited the expression of odontoblast and stem cell markers in the outgrowth area. Three-dimensional explant culture of human dental pulp will provide a novel platform for understanding stem cell biology inside the tooth and developing the regenerative medicine.

• Journal of Plant Biotechnology
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• v.7 no.3
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• pp.155-160
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• 2005

Haloxylon recurvum (Locally known as Khar) is drought and salt tolerant plant of Thar Desert. This plant is a major biomass producer and has economic and ecological importance for the region. There is need for study on biology, propagation and genetic improvement for utilization of this plant for reclamation of saline soils. We report here on in vitro propagation of Haloxylon recurvum (Moq.) using nodal explant. Secretion of phenolic compound from explants was a major constraint for establishment of culture. This was checked by thorough washing and quick transfer of explant on fresh culture medium. Juvenile nodal explant with leaves was found suitable for culture establishment. Benzy-ladenine($4.0\;{\mu}M$) incorporated in Murashige and Skoog (MS) medium with additives (50 mg/L ascorbic acid and 25 mg/L each of adenine sulphate, arginine and citric acid) induced multiple shoots from nodal explant. Addition of $1.0\;{\mu}M$ naphthalene acetic acid (NAA) in combination with $4.0\;{\mu}M$ BAP improved the growth of axillary shoots. Further shoot amplification was achieved by repeated subculture of mother explants on fresh medium. Forty percent of the micropropagated shoots rooted on half-strength MS medium with $4.0\;{\mu}M$ indolebutyric acid (IBA) and 100 mg/L activated charcoal, at $28{\pm}2^{\circ}C$ and $60\%$ RH. Sixty percent of these plantlets were hardened in green house.

• Korean Journal of Plant Resources
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• v.32 no.1
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• pp.19-28
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• 2019

An efficient shoot regeneration condition for pea cv. 'Sparkle' was developed by using optimum explant, plant growth regulator concentrations, and pretreatment of BA onto explant. The average shoot number per explant showed the highest on two kinds of shoot induction media (MSB5 media containing 2 mg/L BA and a combination of 2 mg/L BA and 1 mg/L TDZ) when cotyledonary node explants were cultured. Moreover, the pretreatment of explant in 200 mg/L BA solution was found to be more effective in shoot induction than that of non-pretreatment. By histological study, cell division and proto-meristem were formed near the surface of the sub-epidermal and epidermal cell layers of cotyledonary node in earlier than 3 days after culture. The analysis of genetic stability of regenerants by using thirteen ISSR markers showed that in vitro regenerated plants showed polymorphism with 8.3% compared with their mother plants.

• Journal of Plant Biotechnology
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• v.6 no.2
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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.

• Korean Journal of Medicinal Crop Science
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• v.15 no.1
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• pp.62-66
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• 2007

Plant Regeneration via organogenesis from leaf disk of Korean dandelion was investigated. Leaf disk cultured on MS medium with various combinations of BA (0-4 mg/L) and 2,4-D (0-1 mg/L). Shoot regeneration from leaf explant was observed after 3 weeks of culture. The highest shoot regeneration frequency from leaf disk was obtained with 2 mg/L BA. To analyze the effect of leaf age along shoot formation, we measured number of shoots per explant, shooting rate, fresh and dry weight of leaf explant. The highest number of shoots (11.5) per explant were obtained leaf from 7 weeks old plantlets after seed germination. The regenerated shoots were transferred in 1/2 MS medium with 0.5 mg/L NAA for root formation. Regeneratied plantlets thought organogenesis were growing to whole plants in the pots with acclimation.

• Korean Journal of Plant Tissue Culture
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• v.25 no.5
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• pp.347-351
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• 1998

Plant regeneration via organogenesis from leaf and stipule explants of micropropagated shoots of strawberry (Fragaria $\times$ ananassa cv. Suhong) was achieved. Leaf and stipule explants were detached from shoot-tip cultured shoots and cultured on MS medium with various combinations of BA and NAA under light or dark condition. Shoot regeneration from leaf explant was observed after 3 weeks in culture and was good at the high ratio of BA and NAA among various combination treatments. The highest shoot regeneration frequency from leaf explants was obtained with 1.0 mg/L BA and 0.1 mg/L NAA, in which 31.1% shoot regeneration frequency(1.7 shoots per leaf explant) was yielded. In case of stipule explants, shoot regeneration was largely affected by plant growth regulators during incubation under dark condition for initial 4 weeks but not under continuous light condition. The combination treatment with 2.0 mg/L BA and 0.1 mg/L NAA showed the most excellent shoot regeneration from stipule explants, where 44.4% regeneration frequency(4.0 shoots per explants) obtained. Regenerated shoots were rooted on MS medium with 0.1 mg/L NAA after shoot elongation, and the plantlets regenerated were transferred to soil mixtures with vermiculite and perlite for acclimation.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.29 no.4
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• pp.401-408
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• 1984

This study was conducted to determine the optimum concentrations of growth regulators and their responses on the clonal propagation in axillary bud culture. Cultivars, Hongmi and Shinmi, responded differently to the levels of growth regulators, proliferation rate and shoot growth. The shoot and root of Hongmi cultivar in axillary bud culture were conspicuously induced by combination of NAA(0.1mg/l) and Kinetin(1mg/l) while Shinmi cultivar were affected by the single concentration of Kinetin(1mg/l) and BA(0.1mg/l), and also by the combination of NAA(0.1mg/l) and Kinetin(1mg/l). Better shoot growth and root initiation were obtained in the combination of NAA(0.1mg/l) and Kinetin(1mg/l) regardless of cultivars used when 5mm axillary buds were cultured. The shoots regenerated at the high levels of BA(1-5mg/l) were abnormally thicker and narrower leaves than normal plants and short in shoot height. Frequencies of abnormal plants were higher than that of the low level (0.1mg/l) of BA.