• Title/Summary/Keyword: Tissue-culture

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Chondrogenesis of Mesenchymal Stem Cell Derived form Canine Adipose Tissue

  • Lee, Byung-Joo;Wang, Soo-Geun;Seo, Cheol-Ju;Lee, Jin-Chun;Jung, Jin-Sup;Lee, Ryang-Hwa
    • Proceedings of the KSLP Conference
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    • 2003.11a
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    • pp.183-183
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    • 2003
  • Background and Objectives : Cartilage reconstruction is one of medical issue in otolaryngology. Tissue engineering is presently being utilized in part of cartilage repair. Sources of cells for tissue engineering are chondrocyte from mature cartilage and bone marrow mesenchymal stem cells that are able to differentiate into chondrocyte. Recent studies have shown that adipose tissue have mesenchymal stem cells which can differentiate into adipogenic, chondrogenic myogenic osteogenic cells and neural cell in vitro. In this study, we have examined chondrogenic potential of the canine adipose tissue-derived mesenchymal stem cell(ATSC). Materials and Methods : We harvested canine adipose tissue from inguinal area. ATSCs were enzymatically released from canine adipose tissue. Under appropriate culture conditions, ATSCs were induced to differentiate into the chondrocyte lineages using micromass culture technique. We used immunostain to type II collagen and toluidine blue stain to confirm chondrogenic differentiation of ATSCs. Results : We could isolate ATSCs from canine adipose tissue. ATSCs expressed CD29 and CD44 which are specific surface markers of mesenchymal stem cell. ATSCs differentiated into micromass that has positive response to immunostain of type II collagen and toluidine blue stain. Conclusion : In vitro, ATSCs differentiated into cells that have characteristic cartilage matrix molecules in the presence of lineage-specific induction factors. Adipose tissue may represent an alternative source to bone marrow-derived MSCs.

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Viral Infection of Tissue Cultured Orchids and Evaluation of Damages

  • Chung, Bong-Nam;Yoon, Ju-Yeon;Kim, Mi-Sun
    • The Plant Pathology Journal
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    • v.26 no.2
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    • pp.194-197
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    • 2010
  • Most orchids are propagated by tissue culture. To survey the viral infection of tissue cultured Orchids, total RNA was extracted from in vitro Cymbridium and Phalaenopsis spp. collected from companies producing tissue-cultured orchids, and RT-PCR analysis was conducted with primer pairs specific to Cymbidium mosaic virus (CymMV) and Odontoglossum ring spot virus(ORSV), which are infecting wide range of orchid genera. The bulb size of Cymbidium infected with CymMV and ORSV was compared with healthy one at 10 months after planting in vitro orchids in the glasshouse. The CymMV or ORSV infection in 97 Cymbidium and 55 Phalaenopsis plants was 84.5 and 89.1 %, respectively. Mixed infection was found in 52.6 and 47.3% of Cymbidium and Phalaenopsis tested, whereas virus-free orchids were 15.5 and 10.9%, respectively. The CymMV and ORSV reduced the bulb size by 2.7-50% depending on the cultivars of Cymbidium. The both viruses caused yellowing, mottle and mosaic with or without necrosis in 4 Cymbidium cultivars.

OSTEOGENIC ACTIVITY OF CULTURED HUMAN PERIOSTEAL-DERIVED CELLS IN A THREE DIMENSIONAL POLYDIOXANONE/PLURONIC F127 SCAFFOLD (Polydioxanone/pluronic F127 담체에 유입된 골막기원세포의 조골활성)

  • Lee, Jin-Ho;Oh, Se-Heang;Park, Bong-Wook;Hah, Young-Sool;Kim, Deok-Ryong;Kim, Uk-Kyu;Kim, Jong-Ryoul;Byun, June-Ho
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.31 no.6
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    • pp.478-484
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    • 2009
  • Three-dimensional porous scaffolds play an important role in tissue engineering strategies. They provide a void volume in which vascularization, new tissue formation, and remodeling can occur. Like any grafted materials, the ideal scaffold for bone tissue engineering should be biocompatible without causing an inflammatory response. It should also possess biodegradability, which provides a suitable three-dimensional environment for the cell function together with the capacity for gradual resorption and replacement by host bone tissue. Various scaffolds have already been developed for bone tissue engineering applications, including naturally derived materials, bioceramics, and synthetic polymers. The advantages of biodegradable synthetic polymers include the ability to tailor specific functions. The purpose of this study was to examine the osteogenic activity of periosteal-derived cells in a polydioxanone/pluronic F127 scaffold. Periosteal-derived cells were successfully differentiated into osteoblasts in the polydioxanone/pluronic F127 scaffold. ALP activity showed its peak level at 2 weeks of culture, followed by decreased activity during the culture period. Similar to biochemical data, the level of ALP mRNA in the periosteal-derived cells was also largely elevated at 2 weeks of culture. The level of osteocalcin mRNA was gradually increased during entire culture period. Calcium content was detactable at 1 week and increased in a time-dependent manner up to the entire duration of culture. Our results suggest that polydioxanone/pluronic F127 could be a suitable scaffold of periosteal-derived cells for bone tissue engineering.

Purification and Partial Characterization of a Peroxidase from Perilla Callus

  • Hur, Yeon-Jae;Lee, Han-Gil;Hu, Gaosheng;Chung, Won-Bok;Jeong, Soon-Jae;Yi, Young-Byong;Nam, Jae-Sung;Chung, Young-Soo;Lee, Jai-Heon;Kim, Doh-Hoon
    • Journal of Plant Biotechnology
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    • v.34 no.4
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    • pp.355-361
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    • 2007
  • Cotyledons of perilla6 were cultured on MS medium containing 0.5 mg/l NAA and 0.5 mg/l BA for 7 weeks. The activity of perilla peroxidase was observed to increase following culture stages as assessed by peroxidase assay. A peroxidase (POD) was purified from perilla tissue cultured on MS medium for 7 weeks. The peroxidase was purified using ion exchange and gel nitration chromatography. The perilla peroxidase had a molecular mass of 30 kDa by SDS-PAGE. We showed that the N-terminal amino acid sequence of this protein shared 67% identity with the tea peroxidase. As indicated by SDS-PAGE, the banding pattern of the 30 kDa polypeptide present in total soluble protein from perilla tissue was increased following culture stages. Immunoblot analysis indicated that perilla peroxidase protein appeared after 3 weeks of perilla tissue culture, and continued to increase with extended duration of tissue culture for at least 7 weeks.

Resistance to the Fungal Pathogen Phytophthora infestans of Transgenic Potato Plants Harboring of Chitinase Gene (Chitinase 유전자 도입 형질전환 감자식물체의 역병저항성)

  • Choi, Kyung-Hwa;Yang, Duk-Chun;Kim, Hyun-Soon;Choi, Kyung-Ja;Cho, Kwang-Yeon;Jung, Hyuk
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.3
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    • pp.177-182
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    • 1999
  • A fungal infection assay between normal and transgenic potato harboring chitinase gene in cultivar Belchip was investigated. In the first stage of experiment, seven transgenic lines having 12cm tall were tested for their resistance against potato late blight pathogen Phytophthora infestans by infection with the zoospores, artificially, Susceptibility to potato late blight infection could be classified into three types based on the rate. In terms of resistance to the disease, two lines were higher, two lines were more suppressive, and three lines were similar as compared with the control. In the following experiment, only 2 risistant lines and 1 suppressed line were used to confirm the resistance again. The results of both experiments were similar. Furthermore, two highly resistant transgenic lines grown in field exhibited a higher resistance than control under the conditions of natural ocurrence of the fungal disease.

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Improved Detection and Purification of Grapevine Leafroll-associated 3 Closterovirus Using Tissue Culture (포도 조직배양에 의한 Grapevine Leafroll-associated 3 Closterovirus의 증식과 검출효율 증대)

  • 김현란;정재동;정봉남;이봉춘;박진우;최용문
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.6
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    • pp.335-339
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    • 2001
  • Grapevine leafroll-associated 3 closterovirus (GLRaV-3) is phloem limited virus, and one of the most severe viral diseases found in Korea. However, nonhomogenous distribution and low concentration and seasonal variations of GLRaV-3 in grapevines remain as main problems which prevent the introduction and molecular biology or serology experiments. Virus-infected plantlet in vitro was obtained from node tissue cuttings, which was GLRaV-3 infected 'kyoho' vines. The amount of purified virus was highest in vitro plantlet. Moreover, viruses seem to be relatively homogeneously distributed in all organs including leaf, stem and callus derived from in vitro plantlets. RT-PCR detection using in vitro plantlet tissue as template was most effective. When comparing ELISA to RT-PCR, RT-PCR detection was 1,000 times as effective as ELISA. These results can be explained by improved quality such as tenderness or less tannins in plantlet in vitro. In conclusion, until infected herbaceous host will be available, tissue culture can be usefully adopted as a technique for a good source of GLRaV-3 closterovirus for further studies.

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Saponin Production in Tissue Culture of Ginseng (Panax ginseng C.A. Meyer) (고려인삼의 조직배양에 의한 사포닌 생산)

  • Choi, Kwang-Tae;Park, Ji-Chang;Ahn, In-Ok
    • Journal of Ginseng Research
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    • v.14 no.2
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    • pp.107-111
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    • 1990
  • Ginseng root explants and calli were cultured on modified Murashine and Skoog's media supplemented with different concentrations of organic or inorganic compounds and plant growth requlators to clarify the effects of chemical compositon and plant growth regulators in the medium on the growth of ginseng calli and the production of ginseng saponin. For optimum growth of ginseng calli, the concentrations of 2, 4-D and sucrose were in the range of 1 to 5 mg/l and 1 to 3%, respectively. And it was clarified that sucrose, nitrogen, phosphate, calcium, magnesium, plant growth regulators and their concentrations influcenced the relative biosynthesis of saponin in tissue cultures of Panax ginseng.

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Expression Patterns of CaMV 35S Promoter-GUS in Transgenic Poatoes and Their Clonal Progenies

  • Lee, Kwang-Woong
    • Journal of Plant Biology
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    • v.37 no.1
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    • pp.17-25
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    • 1994
  • Two potato (Solanum tuberosum L.) cultivars were transformed by Agrobacterium tumefaciens harboring cauliflower mosaic virus (CaMV) 35S promoter and $\beta$-glucuronidase (GUS) gene. Expression patterns of the CaMV 35S promoter according to tissue types and developmental stages, and genetic stability of GUS gene were investigated in the clonal progenies of transgenic potatoes. Kanamycin-resistant shoot emerged from tuber disc after 4 weeks of culture, and root was induced 6 weeks after culture on the selection medium. Shooting frequency of cvs. Superior and Dejima were 43% and 27%, respectively. Mature transformants and their clonal progenies showed no phenotypical abnormality. GUS activity was expressed primarily at parenchymatous cells of phloem tissue around the vascular cambium in the stem and root, and higher activity was found at the apical meristem of shoot, root and adventious shoot bud. GUS activity was higher at tubers of young explants than at stored tubers. These facts indicate that expression level of the CaMV 35S promoter differed according to tissue types and developmental stages of the organs. The GUS gene was stably inherited to each clonal progeny and normally expressed.

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Formation and Production of Ascorbate Oxidase by Cucumber Tissue Cultured Cells (오이조직 배양세포에 의한 Ascorbate Oxidase 생성 및 생산)

  • Lee, Jong-Hwa;Chung, Ho-Kwon;Shinmyo, Atsuhiko;Lim, Bun-Sam
    • Microbiology and Biotechnology Letters
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    • v.21 no.4
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    • pp.329-335
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    • 1993
  • Ascorbate oxidase activity in various cucumber tissue extracts was highest in young fruit peeling. Cucumber callus was induced from young fruit peeling and callus cell lines were selected for more than 7 months, which porduced high levels of ascorbate oxidase and had a high growth rate. Induction of callus was optimized with Linsmaier-Skoog(LS) medium at 25$^{\circ}C$ in dark phase. Ascorbate oxidase activity reached a maximum at 5 days after transfer to LS basal liquid-medium ant then declined. The enzyme activity in callus cells was stimulated by addition of 10${\mu}$M $CuSO_4$ in the early logarithmic phase of growth. And also, adding 10${\mu}$M $CuSO_4$ at 3rd day 7th day of culture period, ascorbate oxidase activity in callus cells was maintained to high level. Maximum yield of ascorbate oxidase was found at the 25th day by flask shaking culture, but three-fold of ascorbate oxidase activity was obtained at the 16th day by jar fermentation.

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The Detection of Helicobacter-like Organisms in Dogs (개에서 Helicobacter-like organism의 검출)

  • An, Joong-Ho;Nam, Heon-Woo;Han, Jung-Hee;Kim, Doo
    • Journal of Veterinary Clinics
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    • v.16 no.2
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    • pp.281-288
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    • 1999
  • Helicobacter species have been identified in or isolated from domestic carnivores, but their prevalence in different population of animals and their clinical significance are still unknown. This study was peformed to evaluate the prevalence of Helicobacter in clinically healthy dogs by urease test, culture, morphological examination and polymerase chain reaction (PCR) technique. Tissue samples from 70 dogs in Kangwon and Kyunggi areas from August 1998 to April, 1999, were examined. The detection rates of Helicobacter by urease activity of tissue-samples were 84.6%, 61.3% and 4.8 % in the fundus, the antrum and the duodenum, respectively. One strain of Helicobacter was isolated from the duodenum. It was identified as H canis by biochemical and morphorogical examination. The detection rates of Helicobacter by histological examination were 92.3%, 79.0% and 4.8% in the fundus, antrum and the duodenum, respectively. Helicobacter organisms were colonized more in the gastric pits than in the surface of epithelium, the gastric gland or the parietal cell. Although most of dogs were colonized with Helicobacter in tissue, gross lesions and specific histopathological lesions caused by Helicobacter in these tissues were not observed. The detection rate of Helicobacter by PCR was 78.6%. The histological examination was more sensitive than urease test, culture or PCR technique for the detection of Helicobacter.

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