• Title/Summary/Keyword: In vitro regeneration

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Plant regeneration from suspension-cultured cell clusters of Arabidopsis thaliana (애기장대(Arabidopsis thaliana)의 현탁배양세포괴로부터 식물체 재분화)

  • 김명덕;김준철;진창덕;임창진;한태진
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.3
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    • pp.195-200
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    • 1998
  • Callus induction from leaf and stem explants of Arabidopsis thaliana was successfully obtained when leaf explants were cultured on MS medium containing 2.0 mg/L 2,4-D in the dark and also, when stem explants were cultured on CP medium containing 0.5 mg/L 2,4-D and 0.1 mg/L BAP. Explant-derived sliced calli were suspension-subcultured every week in CP liquid medium with 0.5 mg/L 2,4-D and 0.1 mg/L BAP in the dark, and shoot-forming cell clusters of nodular, pale yellow and knobby type were selected after 7-8 weeks of culture. Shoots were initiated from the green spots of the selected shoot forming calli cultured on MS regeneration medium containing 0.05 mg/L IAA, 7.0 mg/L 2-iP and 30 g/L sucrose under continous illumination for four weeks. Shoot regeneration frequency (calli regenerating at least one shoot) was more than 50%. For plant regeneration, excised shoots were trnasferred to hormone free medium for root initiation after 4 weeks of culture. The regenerants were bolting after 2 weeks of culture and formed in vitro flowering buds within bracts after 4 weeks of culture.

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Biological Effects Of Flurbiprofen Loaded Chitosan To Gingival Fibroblast (Flurbiprofen 함유 키토산 제제가 치은 섬유아세포에 미치는 영향)

  • Chung, Chong-Pyoung;Park, Yoon-Jeong;Lee, Seung-Jin;Rhyu, In-Cheol;Choi, Sang-Mook
    • Journal of Periodontal and Implant Science
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    • v.26 no.1
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    • pp.317-333
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    • 1996
  • The main goal of periodontal regeneration is to be achieved by epithelial exclusion, periodontal ligament cell activation or alveolar bone regeneration. The purpose of this study was to investigate on the physico- chemical and biological characteristics of biodegradable chitosan beads. Chitosan beads were fabricated by ionic gelation with sodium tripolyphosphate and they had the size in 300um diameter. As therapeutic agent, flurbiprofen was incorporated into the beads by 10, 20% loading contents. The release of drugs from the chitosan beads was measured in vitro. Also, biological activity tests of flurbiprofen loaded chitosan beads including cytotoxicity test, ihhibition of $IL-1{\beta}$ production, suppression to $PGE_2$ production, collagenase inhibition test, the ability of total protein synthesis, and tissue response were evaluated. The amount of flurbiprofen released from chitosan was 33-50% during 7 days. Minimal cytotoxicity was observed in chitosan beads. Flurbiprofen released from chitosan beads significantly suppressed the $IL-1{\beta}$ production of monocyte, $PGE_2$ production and markedly inhibited collagenase activity. Meanwhile, flurbiprofen released from this system showed increased ability for protein synthesis. Throughout 4 -week implantation period, no significant inflammatory cell infiltrated around chitosan bead and also fibroblast like cell types at the beads - tissue interface were revealed with gradual degradation of implanted chitosan beads. From these results, it was suggested that flurbiprofen loaded chitosan beads can be effectively useful for biocompatible local delivery system in periodontal regeneration.

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The effects of a combination of calcium sulfate and platelet-derived growth factor on periodontal ligament cells in vitro (Calcium sulfate와 혈소판 유래성장인자의 혼합사용이 치주인대세포에 미치는 영향)

  • Kim, Jun-Seong;Choi, Seong-Ho;Yu, Yun-Jung;Chai, Jung-Kiu;Kim, Chong-Kwan;Cho, Kyoo-Sung
    • Journal of Periodontal and Implant Science
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    • v.27 no.4
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    • pp.785-804
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    • 1997
  • It was well known that calcium sulfate was biocompatible, resorbed rapidly in the body, had potential as a good barrier membrane. Platelet-derived growth factor(PDGF) was one of polypeptide growth factor that had been reported as a biological mediator which regulates activities of wound healing process including the cell proliferation, migration and metabolism. The purpose of this study was to evaluate the effects of a combination of calcium sulfate and PDGF on periodontal ligament cells in vitro to use as a regeneration promoting agent of periodontal tissue. Human periodontal ligament cells were prepared from the premolar tooth extracted for the orthodontic treatment. Cells were cultured in ${\alpha}-MEM$ contained with 20% FBS, at the $37^{\circ}C$, 100% of humidity, 5% $Co_2$ incubator. Cells were inoculated and cultured into 96 well culture plate with $1{\times}10^4cells/well$ of ${\alpha}-MEM$ for 1 day. After discarding the medium, those cells were cultured in ${\alpha}-MEM$ contained with 10% FBS alone(control group), in calcium sulfate(calcium sulfate group), in calcium sulfate treated with 15ng/ml of PDGF-BB(calcium sulfate+PDGF group), in ${\alpha}-MEM$ contained with 10% FBS treated with 15ng/ml of PDGF-BB(PDGF group) for 1, 2, 3 day respectively. And then each group was characterized by examining of the cell counting, MTT assay, collagen synthesis. The results were as follows. 1. In the analysis of cell proliferation by cell counting, both calcium sulfate group and calcium sulfate plus PDGF group showed no stastically significant difference compared to control group, but there was stastically significant difference between PDGF group and calcium sulfate group at 1, 2 day(P<0.05). 2. In the analysis of cell proliferation by MTT assay in calcium sulfate extracts, both calcium sulfate group and calcium sulfate plus PDGF group showed no stastically significant difference compared to control group, but there was stastically significant difference between PDGF group and calcium sulfate group at 2, 3 day, and between calcium sulfate plus PDGF group and calcium sulfate group at 2 day(P

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Preparation and Release Profile of N8f-loaded Polylactide Scaffolds for Tissue Engineered Nerve Regeneration (조직공학적 신경재생을 위한 NGF를 함유한 PLA 담체의 제조 및 방출)

  • 전은경;황혜진;강길선;이일우;이종문
    • Polymer(Korea)
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    • v.25 no.6
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    • pp.893-901
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    • 2001
  • We developed the nerve growth factor (NGF) loaded poly (L - lactide) (PLA) scaffolds by means of emulsion freeze drying method to the possibility for the application of the nerve regeneration of spinal cord disease and the degeneration in Alzheimer's disease. The release amount of NGF from NGF loaded PLA scaffold were analyzed over a 4 week period in vitro at phosphate buffered saline (PBS), pH 7.4, at $37^{\circ}C$. It can be observed the open cell pore structure of porous scaffolds and can be easily controlled the pore structure by the controlling of formulation factors resulting in the controlling of the release rate and the release period. The stability of NGF during the preparation of PLA scaffold was evaluated by comparing the released amounts of total NGF, assayed NGF enzyme - linked immunosorbent assay (ELISA). Released NGF has been found to enhance the neurite sprouting and outgrowth from pheochromocytoma (PC-12) cells. These results suggest that the released NGF from NGF loaded PLA scaffold such as conduit type can be very useful for the nerve regeneration in the neural tissue engineering area.

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Present Status and Prospects of in vitro Production of Secondary Metabolites from Plant sin China

  • Chen, Xian-Ya;Xu, Zhi-Hong
    • Proceedings of the Botanical Society of Korea Conference
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    • 1995.06a
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    • pp.40-56
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    • 1995
  • During the past two decades, China has seen her great progress in plant biotechnology. Since the Chinese market of herb medicine is huge, while the plant resources are shrinking, particular emphasis has been placed in plant tissue and cell cultures of medicinal plants, this includes fast propagation, protoplast isolation and regeneration, cell suspension cultures and large scale fermentation. To optimize culture conditions for producing secondary compounds in vitro, various media, additives and elicitors have been tested. Successful examples of large scale culture for the secondary metabolite biosynthesis are quite limited : Lithospermum ery throrhizon and Arnebia euchroma for shikonin derivatives, Panax ginseng, P. notoginseng, P. quinquefolium for saponins, and a few other medicinal plants. Recent development of genetic transformation systems of plant cells offered a new approach to in vitro production of secondary compounds. Hairy root induction and cultures, by using Ri-plasmid, have been reported from a number of medicinal plant species, such as Artemisia annua that produces little artemisinin in normal cultured cells, and from Glycyrrhiza uralensis. In the coming five years, Chinese scientists will continue their work on large scale cell cultures of a few of selected plant species, including Taxus spp. and A. annua, for the production of secondary metabolites with medicinal interests, one or two groups of scientists will be engaged in molecular cloning of the key enzymes in plant secondary metabolism.

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Plant Regeneration through Micropropagation from Nodal Explants of Critically Endangered and Endemic Plant Exacum travancoricum Bedd

  • Elangomathavan R.;Prakash S.;Kathiravan K.;Seshadri S.;Ignacimuthu S.
    • Journal of Plant Biotechnology
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    • v.8 no.1
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    • pp.51-55
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    • 2006
  • A rapid micropropagation protocol was established for Exacum travancoricum Bedd. The effect of two cytokinins viz. BA and kinetin were studied to evaluate the propagation of plants through nodal explants. MS medium supplemented with 13.32 ${\mu}M$ BA induced early bud break and subsequent production of multiple shoots. Rooting of shoots occurred when cultured on 1/2 strength MS medium supplemented with 14.7 ${\mu}M$ IBA. Rooted plants were acclimatized to greenhouse conditions. The propagated plants were transferred successfully to field with 65% success. As the plant was amenable to propagation in vitro, this can be employed as a tool for conservation of this critically endangered and endemic ornamental herb.

Plant Regeneration through Somatic Embryogenesis from Mature Zygotic Embryos of Ginseng(Panax ginseng C. A. Meyer) and Flowering of Plantlets (인삼(Panax ginseng C. A. Meyer)의 성숙한 배로부터 체세포 배발생을 통한 구분화 및 유식물체의 개화)

  • 이행순
    • Journal of Plant Biology
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    • v.32 no.3
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    • pp.145-150
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    • 1989
  • Mature zygotic embryos dissected from ginseng(Panax ginseng C. A. Meyer) seeds were cultured on Murashige and Skoog's (MS) medium containing various concentrations of 2, 4-dichlorophenoxyacetic acid(2, 4-D) and kinetin. Somatic embryos were induced directly from cotyledonary tissue or from intervening callus. The induction frequency of somatic embryos was up to 55%. Upon transfer to half-strength MS medium supplemented with 1 mg/1 6-benzyladenine(BA) and 1 mg/1 GA3, most somatic embryos developed into plantlets. Over 50% of the plantlets flowered after 4 weeks of culture and then a few bore immature fruits in vitro. Therefore, it is suggested that the juvenility of the ginseng tissue which give rise to somatic embryos does not interfere with in vitro flowering of their regenerated plantlets.

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Effective in Vitro Propagation by Bulb Scale Segments Culture of Muscari comosum var. plumosum

  • Ko Jeong-Ae;Choi Jeong-Ran;Xudong He;Kim Hyun-Soon
    • Korean Journal of Plant Resources
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    • v.19 no.3
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    • pp.432-435
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    • 2006
  • A rapid and mass propagation method for multiple shoots and plant regeneration using bulb scales of Muscari comosum var. plumosum were developed. In vitro different parts of bulb scale as explants were cultured on 11 kinds of MS (1962) media supplemented with various plant growth regulators to induce shoot and callus. A combination of 2.0 mg/L 6-BA and 2.0 mg/L IBA on MS medium was the most favorable and induced the highest production (80%) of shoot formation after 30 days. We also found that the middle part of bulb scale was the best for mass propagation of Muscari comosum var. plumosum of which production could reach 64.4%.