• Title/Summary/Keyword: in vitro cultivation

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Rutin production in hairy root cultures of buckwheat(Fagopyrum esculentum Moench)

  • Park, Sang-Un;Kim, Yeon-Bok;Park, Cheol-Ho
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2002.11b
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    • pp.23-23
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    • 2002
  • We established hairy root cultures of F. esculentum transformed with A. rhizogenes for in vitro rutin production. Additionally, we describe the effects of different media and plant growth regulators on growth and rutin biosynthesis in buckwheat hairy root cultures. Excised leaves of P. tinctorium from 10-day-old seedlings were used as the explant material for co-cultivation with A. rhizogenes 15834. The hairy culture of Fagopyrum esculentum Moench. was established by infecting leaf explants with Agrobacterium rhizogenes 15834. About four to five weeks after co-cultivation with A. rhizogenes, 10 hairy roots were excised from the necrotic explant tissues. After repeated transfer to fresh medium for three months, ten clones were transferred to MS liquid culture medium. The growth and rutin production of each clone differently response to the MS liquid medium. Among these clones, H8, which had exhibited good growth rate and one of the highest rutin productivity, was selected for the following experimment.(중략)

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Studies on the Chemical Control of Mycogone perniciosa Magn in Cultivation of Mushroom Agaricus bisporus(Lang) Sing (양송이 마이코곤병(病)의 방제약제(防除藥劑)에 관한 연구(硏究))

  • Jhune, Chang-Sung;Kim, Gwang-Po;Cha, Dong-Yeul
    • The Korean Journal of Mycology
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    • v.19 no.1
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    • pp.85-87
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    • 1991
  • Prochloraz of fungicide was applied on Mycogone perniciosa causing wet bubble in cultivated mushroom, Agaricus bisporus. In vitro, Prochloraz was an excellent fungicide on two strains of Mycogone, tolerant and non-tolerant to Benomyl, respectively. At the low dosage, Prochloraz more inhibited mycelial growth of mushrooms than Benomyl. At the higher dosage, Benomyl more inhibited the mycelial growth than prochloraz. The higher yield of sporophore of the mushroom with low inferction rate was abtained from several trial of Prochloraz. Prochloroz was concluded to be effective fungicide on Mycogone perniciosa on Agaricus cultivation.

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Antibacterial potential of the extracts derived from leaves and in vitro raised calli of medicinal plants Pterocarpus marsupium Roxb., Clitoria ternatea L., and Sanseveiria cylindrica Bojer ex Hook

  • Shahid, M;Shahzad, A;Anis, M
    • Advances in Traditional Medicine
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    • v.9 no.2
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    • pp.174-181
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    • 2009
  • Pterocarpus marsupium, Clitoria ternatea, and Sanseveiria cylindrica are some of the important and endangered medicinal plant species of India. Despite of medicinal properties, antibacterial potential of the plants have not yet been explored. The present study was designed to optimize the in vitro technique for micropropagation and to screen the extracts from leaves and in vitro raised calli for antibacterial properties. Excised leaf-explants from the parent plants were surface sterilized and cultivated on Murashige & Skoog's (MS) medium containing $N^6$-benzyladenine (BA) in concentrations of 1, 2, 5, and $10{\mu}M$. Optimal growth of calli was noticed at a concentration of $5{\mu}M$, therefore the extracts from calli grown at this concentration were further studied for antibacterial activity. Both alcoholic and aqueous extracts from leaves of respective plants, and their in vitro raised calli were tested for antibacterial activity by agar well diffusion method against a range of Gram-positive and Gram-negative bacteria. Aqueous extracts showed antibacterial activity against limited number of bacterial species; notably the extracts of C. ternatea which showed antibacterial activity against Streptococcus pyogenes, Bacillus subtilis and Bacillus cereus. Alcoholic extracts of all three plants showed antibacterial activity against a wider range of bacteria. Among the Gram-positive bacteria, extracts from C. ternatea showed strong antibacterial activity against Bacillus spp., whereas the extracts of S. cylindrica showed good antibacterial potential for Staphylococcus aureus, S. epidermidis and S. pyogenes. The extracts from all three plants showed antibacterial activity against Gram-negative bacteria, including, Salmonella spp. and Shigella dysenteriae; organisms causing enteric fever and dysentery. In most of the cases, the extracts from respective calli showed comparable, and in some cases better, result in comparison to the extracts from parent leaves. To the best of our knowledge this is the first preliminary report on antibacterial potential, especially through calli extracts, of these plants; and in vitro cultivation of the explants may be used to obtain phytotherapeutic compounds.

Liquid culture of entomopathogenic nematodes

  • Park, Seon-Ho;Yu, Yeon-Su
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.443-444
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    • 2000
  • An in vitro liquid culture media for the cultivation of entomopathogenic nematode Steinernema carpocapsae was developed. Supplementation of whole milk powder with basal liquid culture media showed a remarkable increase in productivity compared to that without whole milk powder and the maximum nematode concentration reached about $1.5{\times}10^5/mL$ within 20 days. Five to twenty gram per liter of liver extract addition revealed highest pathogenicity against 3rd instar of Galleria mellonella which was above about 90% mortality after 48 hr.

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Effect of cell-penetrating peptide-conjugated estrogen-related receptor ${\beta}$ on the development of mouse embryos cultured in vitro

  • Yang, Ning Jie;Seol, Dong-Won;Jo, Junghyun;Jang, Hyun Mee;Yoon, Sook-Young;Lee, Dong Ryul
    • Clinical and Experimental Reproductive Medicine
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    • v.41 no.1
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    • pp.1-8
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    • 2014
  • Objective: Estrogen related receptor ${\beta}$ (Esrrb) is a member of the orphan nuclear receptors and may regulate the expression of pluripotencyrelated genes, such as Oct4 and Nanog. Therefore, in the present study, we have developed a method for delivering exogenous ESRRB recombinant protein into embryos by using cell-penetrating peptide (CPP) conjugation and have analyzed their effect on embryonic development. Methods: Mouse oocytes and embryos were obtained from superovulated mice. The expression of Oct4 mRNA and the cell number of inner cell mass (ICM) in the in vitro-derived and in vivo-derived blastocysts were first analyzed by real time-reverse transcription-polymerase chain reaction and differential staining. Then 8-cell embryos were cultured in KSOM media with or without $2{\mu}g/mL$ CPP-ESRRB protein for 24 to 48 hours, followed by checking their integration into embryos during in vitro culture by Western blot and immunocytochemistry. Results: Expression of Oct4 and the cell number of ICM were lower in the in vitro-derived blastocysts than in the in vivo-derived ones (p<0.05). In the blastocysts derived from the CPP-ESRRB-treated group, expression of Oct4 was greater than in the non-treated groups (p<0.05). Although no difference in embryonic development was observed between the treated and non-treated groups, the cell number of ICM was greater in the CPP-ESRRB-treated group. Conclusion: Treatment of CPP-ESRRB during cultivation could increase embryos' expression of Oct4 and the formation rate of the ICM in the blastocyst. Additionally, an exogenous delivery system of CPP-conjugated protein would be a useful tool for improving embryo culture systems.

A STUDY ON BlOCOMPATABILITY OF RESIN CEMENTS FOR ALL-CERAMIC CROWN (전부도재관용 레진시멘트의 생체적합성에 관한 연구)

  • Kim Kwang-Jun;Kim Sung-Hoon;Jin Tai-Ho
    • The Journal of Korean Academy of Prosthodontics
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    • v.41 no.2
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    • pp.111-127
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    • 2003
  • Statement of problem : Resin cements were used widely on all ceramic crowns, but the influence of resin cements on biocells was not understood clearly. Purpose : This study was investigated to evaluate the biocompatibility of resin cements for all-ceramic crowns. Material and Method : The resin cements used in this study were Panavia F (Kuraray Co., Ltd. Japan), Variolink II (Vivadent Ets., Schann / Liechtenstein), and Bistite II (Bistite dual cure resin cement-clear Tokuyama Soda Co. Japan). The viability of normal human oral keratocytes, gingival fibroblast, and gingival fibroblast immortalized by Human Papilloma virus 16 was measured in vitro for evaluation of cytotoxicity on resin cements, and the response of pulp tissue was analyzed and evaluated with light microscope after application of cements at cutting edge of incisors. Results : The normal human oral keratocytes was the most sensitive to toxicity of resin cement, and toxicity of cements was higher in Bistite II than in Variolink II. The cell viability of immortalized gingival fibroblast did not affected by type of cement and cultivation period, but there was a tendency that cytotoxicity in Bistite II was higher than in Variolink II. The cell viability of gingival fibroblast was similar to that of immortalized gingival fibroblast regardless of cement type, but Bistite II showed more toxic than others after 5 days cultivation. The responses of pulp tissue according to cement type were similar after 2 days cultivation, but revealed high toxicity in Bistite II after 10 days cultivation. Conclusion : Variolink II was more biocompatible than any other resin cements used in this study.

The Effect of Placenta Extract on Proliferation and Differentiation of Human Chondrocytes (태반추출물이 인간 연골세포의 증식과 분화에 미치는 영향)

  • Huh, Jun;Suh, Man Soo;Park, Sae Jung;Lim, Yeung Kook;Shin, Jun Ho;Chung, Ho Yun;Cho, Byung Chae;Park, Jae Woo
    • Archives of Plastic Surgery
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    • v.33 no.5
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    • pp.616-620
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    • 2006
  • Purpose: The isolated human chondrocytes for cartilage reconstruction and transplantation presents a major problem as these cells would change biologically in vitro. For more effective applications of these cells in the clinical field, it is necessary to get a large amount of cells in a short period without affecting their function and phenotype. Methods: This study reports the effects of placenta extract on chondrocytes in vitro. We initiated this study on the basis of the hypothesis that placenta extract can influence both the proliferation of chondrocytes and their biologic functions(for example, to express cell specific gene or to produce their own extracellular matrix). Chondrocytes in monolayer culture with or without placenta extract were collected and analyzed by MTT assay, ECM assay, and RT-PCR. Results: Placenta extract stimulated the proliferation of chondrocytes in monolayer culture. The phenotype of chondrocytes was well maintained during the expansion in monolayers. Chondrocytes expanded in the presence of placenta extract produced ECM, glycosaminoglycan, abundantly. Compared to chondrocyte expanded in culture medium only, chondrocytes expanded with placenta extract demonstrated higher COL2A1 expression that was biochemically comparable to primary chondrocytes. This study provides an evidence that placenta extract is helpful to expand chondrocytes during tissue cultivation, to maintain their differentiated phenotype and to promote their function. Conclusion: These results suggest that placenta extract during cultivation play an important role in controlling cell behaviors. Furthermore, these results provide a biologic basis for cartilage tissue engineering.

Culture characteristics and division process of scuticocilicata in vitro (스쿠티카섬모충의 배양 및 분열)

  • Lee, Chang-Hoon;Ha, Dong-Soo
    • Journal of fish pathology
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    • v.10 no.2
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    • pp.177-186
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    • 1997
  • To establish effectively for isolation and cultivation and cultivation of scuticociliata, we tried to culture scuticociliata using culture medium(P2Y1-DW medium). The number of scuticociliata reached $1.50{\times}10^3$ cells/ml at the temperature of $25^{\circ}C$ in 3 days, but $1.03{\times}10^2$ cells/ml at $15^{\circ}C$. In P1Y1-S medium growth reached $2.6{\times}10^4$cells/ml in 3 days, but the number in DW medium was the lowest with $2.0{\times}10^2$cells/ml. In all these media, the number decreased after 5 days. As the method of the parasite`s division, after attaching the anterior part of the body, its shape gradualy changed to a western pear-like appearance. The parasite completely divided into two cells, then freely swam and became an adult cell, size of $40{\mu}m$ in size within about 3 hours from attaching.

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Purification of IgG1 Type Mouse Monoclonal Antibodies with DEAE-Trisacryl Chromatography (DEAE-Trisacryl 크로마토그래피법에 의한 IgG1 Type 쥐 단일클론 항체의 분리정제)

  • 최태부;정용근
    • Microbiology and Biotechnology Letters
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    • v.16 no.5
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    • pp.335-342
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    • 1988
  • An anion exchange chromatography was employed for the purification of mouse monoclonal antibodies from ascitic fluid and in vitro cultivation media. After cultivation of hybridomas, Alps 25-3, HCGK, A4W, and KW, producing IgG1, the culture supernatants were harvested by centrifugation, precipitated with 50-60% ammonium sulfate, and dialyzed against 0.025 M Tris-HCI buffer (pH 8.2). Then the dialyzed samples were loaded into a DEAE-Trisacryl M anion exchange column. Monoclonal antibodies bound to the DEAE-Trisacryl M were eluted with 0.025 M Tris-HCI buffer (pH 8.2) containing 30-40 mM NaCl. In ammonium sulfate precipitation, the recovery of the monoclonal antibody was shown to be 90% and 84% from in vitro culture media containing 10% and 2% fetal bovine serum, respectively. On the other hand, the pretreatment by ultrafiltration enhanced the yield up to 91% whereas the purity was lower than that by ammonium sulfate treatment. Subsequently, in the DEAE-Trisacryl M chromatographic separation, the purities and recoveries of all the monoclonal antibodies from both the in vitro culture supernatants and ascitic fluids were 70-80% and 65% respectively. The monoclonal antibody, Alps 25-3 could be further purified with a purity of 95% through an immunoadsorbent chromatography.

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In vitro micropropagation of two local taro cultivars for large-scale cultivation

  • Alam, Noor Camellia Noor;Kadir, Abdul Muhaimin Abdul
    • Journal of Plant Biotechnology
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    • v.49 no.2
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    • pp.124-130
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    • 2022
  • The application of traditional taro propagation methods for large-scale cultivation would be insufficient to meet the high demand for quality planting materials. Therefore, this study aimed to develop an in vitro micro-propagation technique for two local taro cultivars (cv.), Wangi and Putih. Taro cormels were collected from the Malaysian Agricultural Research and Development Institute (MARDI) germplasm (Serdang, Malaysia). Explants were taken from the shoot tip of cormels and initially cultured on Murashige and Skoog (MS) basal media for four weeks. The explants were then transferred to different multiplication media supplemented with different types and concentrations of cytokinins such as 6-benzylaminopurine (BAP ) and Thidiazuron (TDZ). Shoot production was quantified after six weeks of culture. The highest mean number of new shoots was produced by the Wangi cultivar on MS medium supplemented with 2.0 mg/l BAP (2.10 shoots), MS medium supplemented with 0.5 mg/l TDZ (2.18 shoots), and Gamborg B5 medium supplemented with 6.0 mg/l BAP (2.43 shoots). The maximum average number of the Putih cultivar shoots was obtained on MS supplemented with 2.0 mg/l BAP (3.57 shoots). MS basal media was used for root initiation, as it produced an average of 25 roots with an 11-cm length. Various types of substrate mixtures were used during acclimatization. The best acclimatization substrate for the Wangi cultivar was 100% peat soil, whereas the Putih cultivar grew optimally in a combination of peat and perlites at a 1:1 ratio. Taro plantlets require approximately 4 to 6 weeks to acclimatize before they can be transferred to the field.