• Title/Summary/Keyword: suspension cell culture

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Determination of the Period of the Formation and Size of Sieve Element Area and Sieve Pore (Streptanthus tortus 조직배양 세포에서 사공의 형성시기와 사공 영역과 사공의 크기 결정)

  • Cho, Bong-Heuy
    • Journal of Plant Biotechnology
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    • v.29 no.1
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    • pp.41-44
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    • 2002
  • During the phloem development from parenchyma cells in a suspension culture of Streptanthus induced sucrose carrier and glucose carrier disappeared. Sieve element area and sieve pore induced suspension culture of Streptanthus were formed almost at the last period of the synthesis of sieve endoplasmic reticulum (SER) and p-protein. The new synthesized cell wall begann to digeste only after the new cell wall was surrounded by SER. The digested region of the cell wall and the formed region of sieve pore were regular comparatively. The completed sieve pore was an oval form, and the outer portion of sieve pore varied, ca 1.2 ${\mu}{\textrm}{m}$~1.6 ${\mu}{\textrm}{m}$ in longitudinal, 0.8 ${\mu}{\textrm}{m}$~1.3 ${\mu}{\textrm}{m}$ in tangential, and the inner size of sieve pore was irregular form of a star-like shape. The number of sieve pore between sieve cells was ca 2~7 per ${\mu}{\textrm}{m}$$^2$ and the sieve pore wall with callose was 0.05 ${\mu}{\textrm}{m}$~0.07 ${\mu}{\textrm}{m}$ in thickness. The energy for the formation of sieve element area and sieve pore might be supplied by mitochondria near the new cell wall and the role of SER remains to be illucidated.

Identification of Water Soluble Metabolites of Pentachlorophenol ( PCP ) in the Suspension Cultures of Soybean and Rice Cells;1. Metabolic Conversion of PCP to Glucose conjugates (대두(大豆) 및 벼 현탁배양(懸濁培養) 중 PCP 수용성대사물(水溶性代謝物)의 동정(同定);1. PCP glucose conjugates의 형성)

  • Kim, Pil-Je;Park, Chang-Kyu
    • Korean Journal of Environmental Agriculture
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    • v.11 no.3
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    • pp.215-223
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    • 1992
  • A metabolic study has been conducted to investigate the conversion of pentachlorophenol(PCP) to water soluble metabolites in soybean and rice cell suspension cultures as well as in intact rice plants. PCP in plant cells was found to be exclusively transformed into water soluble metabolites. The relative rate of the metabolic conversion of PCP in decreasing order was soybean cultures > rice cultures > rice plants. Also observed was that, the older the cultures grown, the lower the conversion rate was. Primary water soluble metabolites isolated from both the 5 day old soybean and 8 day old rice cells were specifically hydrolyzed only by ${\beta}$-glucosidic linkage specific glucosidase, suggesting that the metabolites are ${\beta}$-glucose conjugates. The amount of glucose conjugates was increased with increasing time of incubation of PCP up to 24 hr in both soybean and rice cultures; Thereafter, it was decreased progressively. Most of the glucose conjugates were further metabolized to more polar conjugates in cells, but a portion of them was excreted into the culture medium.

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Intraerythrocytic culture and development of serological diagnostic tests of Babesia gibsoni 2. Intraeryrhrocytic culture of Babesia gibsoni by microaerophilous stationary phase(MASP) (Babesia gibsoni의 적혈구내 배양법과 진단법 개발에 관한 연구 2. Babesia gibsoni의 적혈구내 배양)

  • Suh, Myung-deuk;Joo, Bo-hyun
    • Korean Journal of Veterinary Research
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    • v.38 no.2
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    • pp.359-365
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    • 1998
  • This study was conducted to isolate the protozoan parasite Babesia gibsoni by intraerythrocytic culture method of micoraerophilous stationary phase(MASP) and evaluate the possibility of application for the detection of B gibsoni in canine babesiosis. Also, indirect fluorescent antibody test(IFAT) and thick blood smear(giemsa stain), direct light microscopy (DLM), as control diagnostic tests, were conducted to compare diagnostic effects between MASP, IFAT and DLM. The results obtained from this study were summarized as follows. The protozoan parasite B gibsoni multiplied in 24-well polystyrene plate containing 1.2ml of canine red blood cell suspension in RPMI 1640 medium(pH 7.0) which is contained 20~40% normal canine serum(NCS) under the MASP condition of 5% $CO_2$ and 95% air at $37^{\circ}C$ incubator. Under the above MASP culturing system the percentage of parasitized erythrocytes(PPE) after incubation for 9 days reached the peak. The levels of PPE in MASP culture were shown more higher by exchanging the medium at 24 hour intervals. The parasite were purely isolated from MASP culture of canine red blood cells collected from dogs(pit bullterrier) infected with B gibsoni naturally. Among the total of 83 heads of pit bullterrier blood samples the positive rate was 32 heads(38.5%) in DLM, 45 heads(54.2%) in IFAT and 42 heads(50.6) in MASP culture. In negative cases of IFAT and DLM the isolation rates of B gibsoni by MASP culture were 16 heads(42.1%) of 38 heads and 16 heads(28.6%)% of 56 heads, respectively. From this study it was suggested that MASP culture method by RPMI 1640 medium was a reliable and useful diagnostic test for the diagnosis of B gibsoni infections in canine babesiosis.

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The Growth of Proteolytic Bacteria Immobilized in Capsule Type

  • Han Bong-Ho;Choi Su-Il;Kim Seong-Bong;Kim Sang-Ho
    • Fisheries and Aquatic Sciences
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    • v.2 no.1
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    • pp.44-51
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    • 1999
  • Proteolytic bacteria isolated from fermented anchovy jeotkal were immobilized in capsule type with $0.8\%$ sodium alginate and $CaCl_2/carboxymethyl$ cellulose (CMC). For making the immobilized capsule, the optimal concentration of both $CaCl_2$ and CMC, with respect to the membrane hardness and the growth of proteolytic bacteria in capsule, were $2.0\%$ at following conditions: flow rate of $CaCl_2/CMC$ solution and cell suspension were respectively 3.54 ml/min and 0.15 ml/min when inside diameter of inner and outer capillary tube in immobilizing apparatus were 0.32mm, 0.74mm, respectively. The density of proteolytic bacteria in capsule reached maximum, i.e. $10^8-10^9cells$/capsule during culture under optimal conditions in TPY broth, and these were $10^2-10^4$ times higher than these of before culture. During culture of proteolytic bacteria immobilized in capsule type (PBImC) for 72hrs, few growing cells were lost in the outer medium.

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Inhibitory Effects of Bacterial Isolate Stenotrophomonas sp. KTGBP10 against Viral Infection to Tobacco Plants (세균 Stenotrophomonas sp. KTGBP10의 식물 바이러스 감염억제효과)

  • Kim Young-Sook;Hwang Eui-Ii;Oh Jung-Hoon;Kim Kab-Sig;Yeo Woon-Hyung
    • Journal of the Korean Society of Tobacco Science
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    • v.26 no.2 s.52
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    • pp.79-84
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    • 2004
  • During the screening of antiviral substances having inhibitory effects on tobacco mosaic virus (TMV) infection to tobacco plants, we found a bacterial isolate KTGBP10, which was identified as a Stenotrophomonas sp., strongly inhibited the infection of TMV. When the culture filtrate from KTGBP10 was applied on the upper surface of leaves of Xanthi-nc tobacco plants at the same time or 24 hours before TMV inoculation, almost complete inhibition of TMV infection was achieved. And $40\%$ inhibition was shown with application of the culture filtrate to the under surface of leaves. In field trials, transmission of TMV from diseased seedlings to the healthy ones during transplanting work was reduced by $87.1\~92.6\%$ when the culture filtrate or cell suspension was sprayed onto the tobacco seedlings, cv. NC82, 24 hours before transplanting. No toxic effect was observed on the tobacco plants. When the broth filtrate of KTGBP10 was supplied by soaking through the cut-leaves before and/or after virus inoculation, the TMV infection was also inhibited by $50.4\~65.3\%$.

Mass Production of Eleutherococcus senticosus Plants through in vitro Cell Culture (세포 배양을 통한 가시오갈피(Eleutherococcus senticosus) 묘목의 대량 생산 시스템 개발)

  • Han, Jeong-Yeon;Choi, Yong-Eui
    • Journal of Plant Biotechnology
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    • v.30 no.2
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    • pp.167-172
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    • 2003
  • Immature zygotic embryos of Eleutherococcus senticosus seeds matured rapidly within one month when the seeds comprising zygotic embryos were pieced to small size and cultured on 1/2 MS medium. Frequency of somatic embryos formation was declined rapidly when the zygotic embryos germinated and grew to plantlets. Embryogenic cells were induced by consecutive subculture of somatic embryos on MS medium with 1.0mg/L2,4-D. After heart-shaped somatic embryos were induced by suspension culture, these embryos were plated onto petri dish to support maturation of embryos. Germination of embryos occurred on medium with 5mg/L GA$_3$and transferred to culture bowl to stimulate the further growth. Frequency of soil survival of plantlets was influenced by soil mixture (perlite and peatmoss). The suitable combination of perlite and peatmoss was 1:5, and the soil survival rate was 78% after 4 months. The soil transferred plantlets were over-wintered in field condition after defoliation. New year sprouting of plants was achieved successfully and they grew to adult plants. These results indicate that the systematic procecure of plant production in E. senticosus for micro propagation.

이상추출배양을 통한 Botryococcu braunii에서의 탄화수소 생산

  • Sim, Sang-Jun;An, Jin-Yeong;Kim, Byeong-U
    • 한국생물공학회:학술대회논문집
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    • 2002.04a
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    • pp.112-115
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    • 2002
  • The green colonial algae Botryococcus braunii is characterized by unusual high hydrocarbon contents, ranging from 15 to 75% of dry weight, as long-chain unsaturated hydrocarbons. In two-phase bubble column using various organic solvents, poor recovery 08 - 32%) of hydrocarbon seems to be caused by insufficient mixing between two phases, which was operated using only aeration on the narrow interface between hydrophobic solvent and cell suspension. In addition, hydrocarbon was entrapped tightly in cell-matrix (formed by exopolysaccharide) of algal colony, which make difficult to extract using two-phase system. To improve recovery efficiency, mixed-solvent of extractive solvent (octane) and biocompatible solvent (octane) was tested in two-phase column for in situ extraction. In two-phase extraction culture using mixed-solvent, the algal growth was intensely inhibited even at low concentration of polar octanol solvent. the hydrocarbon recovery in two-stage cell-recycle extraction showed a 2.9 fold increase (57%) over that in two-phase extraction. Up to 60 % of hydrocarbon could be recovered without serious cell-damage in the case of downstream separation for 6 h at the high recycle flow rate using this process after batch culture.

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A study of the effects of PDGF-BB on the characteristics of bone stromal and periodontal ligament cells (혈소판유래성장인자-BB가 골간질세포와 치주인대세포의 성상에 미치는 영향)

  • Kwon, Young-Hyuk;Park, Joon-Bong
    • Journal of Periodontal and Implant Science
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    • v.26 no.4
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    • pp.949-965
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    • 1996
  • The main goal of periodontal therapy is to restore the lost periodontal tissue and establish the attachment appratus. Current acceptable therapeutic techniques are included : removal of diseased soft tissue, demineralization of exposed root surface, using the barrier membrane for preventing the downgrowth of gingival epithelial cell, insertion of graft materials as a scaffolding action, and biological mediators for promoting the cell activity. The latest concept one among them has been studied which based on the knowledge of cellular biology of destructed tissue. Platelet-derived growth factor(PDGF) is one of the polypeptide growth factor which have been reported as a biological mediator to regulate activities of wound healing progress including cell proliferation, migration, and metabolism. The purposes of this study is to evaluate the influences of the PDGF as biological mediator to periodontal ligament and bone marrow cell. Both right and left maxillary first molar were extracted from rat which had treated with 0.4% ${\beta}-Aminopropionitril$ for 5 days, and feeded until designed date to sacrifice under anesthesisa. Periodontal ligament were removed from the extracted socket of the rat, and cultured with Dulbecco's Modified Essential Medium(DMEM) contained with 10% Fetal Bovine Serum, 100U/ml penicillin, $100{\mu}g/ml$ streptomycin, $0.5{\mu}g/ml$ amphotericin-B. Bone marrow cell were culture from bone marrow suspension with which washed out from femur with same medium. The study was performed to evaluate the effect of PDGF to periodontal ligament and bone cell, cell proliferation rate, total protein synthesis, and alkaline phosphatase activity of rat periodontal ligament(PDL) cell and bone stromal(RBS) cell in vitro. The effects of growth factors on both cells were measured at 3, 5th day after cell culture with (control group) or without growth factors(experimental group). The results were as follows: 1. The tendency of cell proliferation under the influence of PDGF showed more rapid proliferation pattern than control at 3 and 5 days after inoculation. 2. The activity of Alkaline phosphatase revealed 14, 80% increased respectively at 3, 5 days culture than control group. Measurements of ALPase levels indicated that PDL cells had significantly higher activity when compared with that of co-culture groups and GF only(P<0.05). And, ALPase activity in 10 days was higher than that of 7 days(P<0.05). 3. The tendency of formation of the mineralized nodule were observed dose-depend pattern of PDL cells. There was statistically significant difference among group l(PDL 100%), 2(PDL 70%:GF 30%), and 3(PDL 50%:GF 50%)(P<0.01). But, there was no difference among group 3, 4(PDL 30%:GF 70%), and 5(GF 100%). 4. Also, the number of nodule was greater in co-culture of PDL 70% and GF 30% than in culture of PDL 70%(P<0.05). From the above results, it is assumed that the PDGF on PDL cells and RMB cell culture. GF stimulates the cell growth, which is not that of PDL cells but GF. And, the activity of ALPase depends on the ratio of PDL cells, and ALPase may relate to the initial phase of nodule formation. Also, it is thought that the calcified nodule formation principally depends on PDL cells, is inhibited by GF, and affected by cell density. In conclusion, platelet-derived growth factor can promote rapid osteogenesis during early stage of periodontal tissue regeneration.

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Maintenance and Differentiation of Pluripotential Embryonic Cell Lines from Mouse Blastocysts (BCF1 생쥐 배반포기 유래 배아간세포 작성에 관한 연구)

  • 이재원;이훈택;정길생
    • Korean Journal of Animal Reproduction
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    • v.18 no.4
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    • pp.235-244
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    • 1995
  • The present study was designed to demonstrate that ES cell lines efficiently could be isolated from explanted blastocysts of hybrid BCF1 mouse when grown on STO feeder layer derived from mouse fibroblasts in culture medium supplemented with leukemia inhibitory factor (LIF). The expanded blastocysts were attached to mitomycin C-inactivated STO feeder layer and were cultured for 4 days. Four days later the ICM was disaggregated by a short term trypsin treatment (0.25% trypsin / 0.04% EDT A for 2-3 min). The resulting cell suspension was seeded on a new STO feeder layer and covered with DMEM supplemented with 10% FCS, 0.1 mM nonessential amino acid, 0.1 mM sodium pyruvate, 0.1 mM mercaptoethanol and 1,000 U/ml LIF. Colonies of ES-like cells were observed after the second passage. These colonies were repeatedly passaged at approximately 5 day intervals. In this study, five ES-like celllines were isolated by directly explanting blastocysts, but three lines were lost after the 5th passage, possibly due to toxic effects of a new FCS batch. The characterization of developmental potential of isolated cell lines was performed with respect to in vitro differentiation and specific activity of alkaline phosphatase (AP). When cells were cultured in suspension, the aggregates of cell lines were capable of forming simple embryoid bodies (EB), and showed the capacity for forming cystic multilayer EBs. In addition, the cell lines were positive for AP staining, a biochemical marker characteristic of mouse ES cells.

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Developmental and Structural Diversity of Regenerated Plants in Cell and Tissue Cultures (세포조직배양계에서 재생된 식물의 발생 및 형태학적 다양성)

  • 소웅영
    • Proceedings of the Botanical Society of Korea Conference
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    • 1993.07a
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    • pp.1-36
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    • 1993
  • It is possible to regenerate plants from calli, single cells and protoplasts of numerous species via organogenasis or embryogenesis in cell and tissue culture systems. Also such regeneration of plants can directly occur from cells of explants. However certain plant species has not been yet provided cultures suitable for plant regeneration from cells or tissues. For example, we have to confirm the regenerability of plant from cells before preparing transformed cells for application. Even more, it is very important to notice that regenerated plants in cell and tissue cultures often show structural abnormality. The mojority of those plants is functionally disordered and eventually cases degenerated. One of such examples is vitreous plants which are manifested mainly in the leaves and manifesteds to a lesser extent in the stems and roots. Regenerants in suspension cultures show more frequent vitrification than on gelled media so that relative humidity and water potential are the key factors involved in abnormal morphogenesis in vitro. The other is that somatic embryos formed in media containing BAP or high concentration of sucrose show frequently cotyledon aberrancy such as polycotyledon and born type cotyledon. The embryos with aberrant cotyledon of Codonopsis lanceolata could not germinate or regenerate into plants in many cases. In contrast, the polycotyledon embryos of Aralia cordata germinated in higher percentage than two cotyledonary embryos, but horn type cotyledonary embryos rarely germinated. The major cause of poor germination is the abnormal development of plumule apex meristem.

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