• 제목/요약/키워드: tissue cultures

Search Result 325, Processing Time 0.027 seconds

Characterization of Anionic Peroxidase Induced by Low Host-Specific Elicitor in Suspension Cultures of Rose (Rosa sp.) (장미(Rosa sp.) 현탁배양세포에서 숙주 특이성이 낮은 Elicitor에 의해 유도되는 Anionic Peroxidase의 특성)

  • 신미선;양은진;이인철
    • Korean Journal of Plant Tissue Culture
    • /
    • v.25 no.4
    • /
    • pp.277-282
    • /
    • 1998
  • Whereas cationic extracellular peroxidases (PODs) were observed in the suspension cultures of rose (Rosa sp. L. cv Pual's scarlet) grown under normal conditions, new anionic isozymes were induced within 24 hr by the treatment of low host-specific elicitor (10 mg glucan/L media) prepared from yeast cell wall. Prominent anionic (pI 6.1) and cationic POD (pI 8.4) were purified and characterized to understand the physiological role of the enzymes. Both enzymes were purified (ca.200 fold) by the ammonium sulfate precipitation, ion exchange chromate-graphy and gel filtration chromatography. The Km values of the purified anionic POD for ferulic acid and $\textrm{H}_2\textrm{O}_2$ were 4.64 mM and 0.72 mM, whereas those of the cationic POD were 1.38 mM and 0.48 mM, respetively. The activity of the anionic POD as NADH oxidase was twice higher than that of cationic POD. The NADH oxidation in the anionic POD fraction was inhibited by 60% on the addition of 0.1 mM coniferyl alcohol, while that in the cationic fraction was inhibited by 15%.

  • PDF

Effects of Excising In Vitro-Formed Roots on Acclimatization of Micropropagated Cassava Plantlets (카사바의 미세증식에서 기내 발생 부정근의 절단이 순화에 미치는 영향)

  • Yoon, Sil;Cho, Duck-Yee;Soh, Woong Young
    • Korean Journal of Plant Tissue Culture
    • /
    • v.28 no.2
    • /
    • pp.103-108
    • /
    • 2001
  • The in vitro plantlets of cassava (Manihot esculenta Crantz cv. MColl 22) could be regenerated from nodal explant cultures in a liquid MS basal medium containing 0.01 mg/L zeatin for 2 weeks. The plantlets of 1.5∼2.5 cm in shoot length were transplanted to a glass bottle containing fine sand and acclimated under non-sterile conditions after excising their intact roots by: 1) prune leaving roots base of 1∼1.5 cm; 2) complete removal of roots; and 3) cutting off the rooting zone. The majority of in vitro-formed intact roots continued growth after transferred to soil, and all of the damaged roots stopped further growth. The plantlets with excised roots began to develop new roots within 7∼10 days after being transferred to a glass bottle, and a few of the pruned roots developed lateral roots from the remaining portion. Pruning and removal of in vitro roots resulted in a high survival rate (>87%), and did not significantly affect ex vitro root regeneration and acclimation, but the plantlets in which the rooting zone had been cut-off showed 73% survival rate. Pruning or removal of in vitro roots before transfer of plantlets is recommended for useful method of commercial micropropagation because of easier handling and high survival rate of plantlets.

  • PDF

Effect of exogeneous plant growth regulators on morphogenetic response in vitro by embryo and leaf cultures of Camellia sinensis(L.) O. Kuntze (차나무 잎과 배 배양에 있어서 식물 생장조절물질이 형태형성에 미치는 영향)

  • PARK, Young Goo;AHN, In-Suk;BOZHKOV Peter
    • Korean Journal of Plant Tissue Culture
    • /
    • v.24 no.3
    • /
    • pp.129-135
    • /
    • 1997
  • Morphogenetic responses were investigated by culturing embryo and leaf explants of Korean wild type tea plant, Camellia sinensis (L.) O. Kuntze. Induction of direct somatic embryogenesis as well as adventitious and/or axillary shoots was obtained from mature zygotic embryo cultures on Murashige and Skoog (MS) basal medium having 5 to $20\mu\textrm{M}$cytokinin a lone. Morphogenetic response was decreased dramatically by the addition of auxins tested. One hundred percent of induced and isolated shoots formed roots after four weeks of culture on half-strength MS or quarter-strength Schenk and Hildebrandt (SH) media supplemented with $10\mu\textrm{M}$indole-3-butyric acid (IBA). Immature zygotic embryos were shown to be a suitable explant for embryogenic callus formation in the presence of 2, 4-dichlorophenoxyacetic acid(2, 4-D) in basal medium. Mature zygotic embryo originated leaves were used to test their ability for mophogenesis by incorporating plant growth regulators such as IBA, naphthyl-1-acetic acid (NAA), and 6-benzylaminopurine (BAP). Apparently, the morphogenetic responses of the cultured explant sources on the types and/or levels of plant growth regulators tested were observed visually.

  • PDF

Practical Propagation Methods for Production of Prothalli and Sporophytes in Deparia pycnosora (Christ) M. Kato

  • Jang, Bo Kook;Park, Kyungtae;Cho, Ju Sung;Lee, Cheol Hee
    • Proceedings of the Plant Resources Society of Korea Conference
    • /
    • 2019.04a
    • /
    • pp.43-43
    • /
    • 2019
  • Deparia pycnosora (Christ) M. Kato is a fern used as ornamental plant. In addition, it is called "Teol-go-sa-ri" in Korean name. The aim of this study was to develop a practical propagation method of D. pycnosora using tissue culture technique. Prothallus obtained from spore germination was the used as experiment materials. The prothalli (300 mg) used in all experiments were sub-cultured for 8-week intervals. The most suitable media for prothallus propagation were identified by culturing 300 mg of prothalli in $1/4{\times}$, $1/2{\times}$, $1{\times}$, $2{\times}$ MS medium and in Knop medium for 8 weeks. Also, the prothalli were cultured by chopping with a scalpel. In addition, sucrose, activated charcoal, and total nitrogen source were added in different concentrations based on the culture medium selected. Cultures were maintained at a temperature of $25{\pm}1^{\circ}C$, light intensity of $30{\times}1.0{\mu}mol-m-2{\cdot}s-1$, and a photoperiod of 16/8 h (light/dark) in in vitro. The results showed that optimum was achieved prothallus fresh weight and development in $1{\times}$ MS medium. When other components were added to the basic $1{\times}$ MS medium, prothallus propagation was maximized in $1{\times}$ MS medium supplemented with 2% sucrose, 0.2% activated charcoal, and 60 mM total nitrogen. To select a suitable soil mixture for sporophyte formation, 1.0 g of prothallus was blended with distilled water, spread on five combinations of different soil substrates (decomposed granite, horticultural substrates, peat moss, and perlite), and cultivated for 12 weeks. The sporophyte cultures were maintained at a temperature of $25{\pm}1^{\circ}C$, light intensity of $43{\pm}2.0{\mu}mol-m-2{\cdot}s-1$, humidity of $84{\pm}1.4%$, and a photoperiod of 16/8 h (light/dark). As a results, horticultural substrate alone, 2:1 (v:v) mixtures of horticultural substrate and perlite, and 2:1 mixtures of horticultural substrate and decomposed granite induced 208.0, 201.3 and 248.8 sporophytes per pot, respectively. Therefore, this result could provide a practical mass propagation method of D. pycnosora

  • PDF

Relationship with Passage Time of Human Dental Pulp Stem Cells from Supernumerary Tooth by Classification (과잉치 분류에 따른 치수유래줄기세포 계대 배양 시간의 연관성)

  • Shin, Yeoseob;Kim, Jongbin;Kim, Jongsoo
    • Journal of the korean academy of Pediatric Dentistry
    • /
    • v.43 no.4
    • /
    • pp.419-426
    • /
    • 2016
  • For this research 20 supernumerary teeth impacted in the maxillary anterior have been extracted and pulp cells have been collected from them. From the collected pulp cells, total of 17 (10 males, 7 females) have been selected as subjects. From this research, the run-time of successive culture of the cell from tooth number pulp tissue was $2.91{\pm}0.29$ days. From the gathering of cells from the initial pulp tissue until gaining 80% confluency took $4.53{\pm}0.94$ which was the longest. The following successive cultures took $2.73{\pm}0.32$ days. Average runtime for female was $2.81{\pm}0.27$ days whereas male had average runtime of $2.98{\pm}0.29$ days. Average run-time for inversion was $2.94{\pm}0.30$ days and for normal location, $2.80{\pm}0.20$ days. Average runtime was $2.92{\pm}0.31$ days and other forms took $2.88{\pm}0.22$ days. In the future, follow up research would be needed to evaluate the efficiency of the cells collected from the initial passage and the latter passage as stem-cells and taking into consideration the less than 3 days'time for the subculture, it could be concluded that the research efficiency and fast cultivation would be sufficiently effective.

Immunomodulatory Effect of Mesenchymal Stem Cell-Derived Exosomes in Lipopolysaccharide-Stimulated RAW 264.7 Cells (Lipopolysaccharide로 자극한 RAW 264.7 세포에서 성체줄기세포 유래 엑소좀(exosome)의 면역 조절 효과)

  • Jung, Soo-Kyung;Park, Mi Jeong;Lee, Jienny;Byeon, Jeong Su;Gu, Na-Yeon;Cho, In-Soo;Cha, Sang-Ho
    • Microbiology and Biotechnology Letters
    • /
    • v.44 no.3
    • /
    • pp.383-390
    • /
    • 2016
  • Mesenchymal stem cells (MSCs) are multipotent stem cells that can be differentiated into a variety of cell types, including adipocytes, osteoblasts, chondrocytes, β-pancreatic islet cells, and neuronal cells. MSCs have been reported to exhibit immunomodulatory effects in many diseases. Many studies have reported that MSCs have distinct roles in modulating inflammatory and immune responses by releasing bioactive molecules. Exosomes are cell-derived vesicles present in biological fluids, including the blood, urine, and cultured medium of cell cultures. In this study, we investigated the immunomodulatory effects of mouse adipose tissue-derived MSCs (mAD-MSCs), cultured medium (MSC-CM) of mAD-MSCs, and mAD-MSC-derived exosomes (MSC-Exo) on lipopolysaccharide (LPS)-induced RAW 264.7 cells. We observed that the expression levels of IL-1β, TNF-α, and IL-10 were significantly increased in LPS-stimulated RAW 264.7 cells compared to those in LPS-unstimulated RAW 264.7 cells. Additionally, these values were significantly (p < 0.05) decreased in mAD-MSCs-RAW 264.7 cell co-culture groups, MSC-CM-treated groups, and MSC-Exo-treated groups. MSCs can modulate the immune system in part by secreting cytokines and growth factors. We observed that immunomodulatory factors such as IL-1β, TNF-α, and IL-10 were secreted by mAD-MSCs under co-culturing conditions of mAD-MSCs with activated RAW 264.7 cells. In addition, mAD-MSC-derived exosomes exhibited similar immunomodulatory effects in activated RAW 264.7 cells. Therefore, our results suggest that mAD-MSCs have an immunomodulatory function through indirect contact.

Development of a Primary Tissue Culture Method having Greater Reliability than Isolated Cell Cultures - Steroid-Responsiveness of Uterine Myometrial and Myomatous(Leiomyomatous) Cells (자궁근종세포의 최적 초기배양 조건 확립 - 정상 자궁근세포와 자궁근종세포의 스테로이드에 대한 반응)

  • Lee, Eun-Ju;Bajracharya, Prati;Hyun, Jin-Hee;Kim, Hang-Jin;Song, Gun-Ho;Cho, Kyung-Hyun;Lee, Dong-Mok;Lee, Taek-Hoo;Chun, Sang-Sik;Choi, In-Ho
    • Development and Reproduction
    • /
    • v.11 no.3
    • /
    • pp.205-217
    • /
    • 2007
  • As an initial step toward better understanding of the molecular mechanism of estrogen-dependent growth in myoma, an optimal primary cell culture condition has been developed and examined by this study. Myoma and myometrium were cultured by two different methods. Culture stability and $E_2$-responsiveness in stable culture were studied. The culture of digested tissue pieces(Method 2) was found to be a stable culture method for the myoma and myometrium showing a favorable response to estrogen. mRNA expression of PR, IGF-1 and IGF-1 receptor genes was enhanced by $E_2$. The gene responses to $E_2$ were higher in myoma compared with myometrium. Moreover, these responses were more expressive in tissues than in the surrounding cells in primary culture of normal myometrium and myoma, implying a vital role of cell communication through the extracellular matrix in maintaining the estrogen-responsiveness. The development of an improved cell culture system for myoma provides an in vitro tool to further investigate the basis of the tumor formation.

  • PDF

Detection for Methicillin Resistant Staphylococcus aureus in Using Bio-Chip Based Loop Mediated Isothermal Amplification Assay (칩 기반 등온증폭법을 이용한 약제 내성 포도상구균의 검출)

  • Cho, Min-Ho;Jang, Won-Cheoul;Choi, Jae-Gu
    • Journal of the Korean Chemical Society
    • /
    • v.57 no.1
    • /
    • pp.81-87
    • /
    • 2013
  • Staphylococcus aureus is the most important pathogen in nosocomial infections, including bloodstream infections. Prompt identification of S. aureus from blood cultures and detection of methicillin resistance are essential in cases of suspected sepsis. We have studied a new method for the sequence-specific visual detection of minute amounts of nucleic acids using intercalating reaction by addition of SYBR Green to amplicons of LAMP, and it's a unique gene amplification method in which DNA can be isothermally amplified using only one enzyme. Staphylococcus-LAMP, which targets the spa gene, encoding S.aureus-specific protein A, and the mecA gene, encoding penicillin-binding protein-2' for methicillin resistance, detected MRSA and MRSE. In this study, by using LAMP assay, I detected for Staphylococcus aureus and Staphylococcus epidermidis concentration in the clinical sample. The detection of Staphylococcus aureus and Staphylococcus epidermidis was tested by using serial 10-fold dilutions standard solution. I have accurate detected the limit of detection, sensitity, specificity and reproducibility of the assay. The Bio-chip based LAMP assay allowed easy, rapid, accurate and sensitive detection of infection with Staphylococcus and especially applicable in a resource-limited situation.

Light Influences, Morphogenesis and Protein Content on Callus Differentiation of Cucumber (Cucumis sativus L.) (오이 캘러스 분화시 광의 영향, 형태발생 및 단백질함량)

  • 이은모;조만현;송남현;우인식;이영복;곽상수
    • Korean Journal of Plant Tissue Culture
    • /
    • v.27 no.3
    • /
    • pp.213-217
    • /
    • 2000
  • To investigate the cucumber regeneration from embryogenic calli, shoot tips of aseptically-grown cucumber seedlings were used as explants for establishing tissue cultures. Growth and differentiation of callus were studied by using Murashige and Skoog's (MS) medium containing 0.5 to 2 mg/L 2,4-D. Plantlets were induced from shoot tip culture on the plant growth regulators-free MS medium. Non-embryogenic calli and viscous calli were induced on the medium supplemented with 0.5 to 2 mg/L 2,4-D, but embryogenic callus was not induced on the same medium. Segments (ca. 5∼10 mm) of aseptically-grown hypocotyl from five to seven days old seedlings after germination were placed on MS medium supplemented with 1 mg/L 2,4-D for 50 days. Embryogenic calli and embryoids were induced only from the seedlings grown in dark condition, and hypocotyl was placed on the media explanted in light condition. Foully-five point one percent of white fragile calli and 0.6% yellowish compact calli formed roots. Yellowish callus lines were investigated to have a considerably higher concentration of crude proteins than white callus lines. Plantlets derived from embryogenic calli or embryoids have been transferred to pots containing sterile vermiculite and perlite. Normal fruits were harvested from nutrient culture on aggregated hydroponics in the F-clean house.

  • PDF

Effect of Plant Growth Regulators on Direct Shoots Formation and Somatic Embryogenesis from Leaf Tissue Culture of Muscari armeniacum 'Early Giant' (무스카리(Muscari armeniacum 'Early Giant') 엽절편 조직으로부터 신초형성과 체세포 배발생에 미치는 생장조절물질의 영향)

  • Jeon, Su-Min;Chung, Mi-Young;Lee, Hyang-Bun;Han, Jeung-Sul;Park, Jae Suk;Kim, Chang-Kil;Chung, Jae-Dong
    • FLOWER RESEARCH JOURNAL
    • /
    • v.18 no.4
    • /
    • pp.261-265
    • /
    • 2010
  • This experiment was carried out in order to determine proper plant growth regulators (PGR) and their concentrations for direct shoot induction and somatic embryogenesis from leaf tissue cultures of Muscari armeniacum 'Early Giant'. Direct shoot formation from the leaf explant culture was effective only on a half-strength MS medium containing $0.1mg{\cdot}L^{-1}$ 2,4-D, while embryogenesis was occurred on a half-strength MS medium containing $0.1{\sim}1.0mg{\cdot}L^{-1}$ IPA or without PGR. The regenerated bulblets derived from embryos or shoots were harvested and transplanted into a greenhouse. The sprouting percentage of bulblets obtained from different culture media ranged from 80 to 100% and growth of quality bulblets was enhanced when the bulblets were harvested from the medium containing $0.1mg{\cdot}L^{-1}$ NAA and $1.0{\sim}3.0mg{\cdot}L^{-1}$ IPA.