• Title/Summary/Keyword: tissue cultures

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Differentiation and characteristics of undifferentiated mesenchymal stem cells originating from adult premolar periodontal ligaments

  • Kim, Seong Sik;Kwon, Dae-Woo;Im, Insook;Kim, Yong-Deok;Hwang, Dae-Seok;Holliday, L. Shannon;Donatelli, Richard E.;Son, Woo-Sung;Jun, Eun-Sook
    • The korean journal of orthodontics
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    • v.42 no.6
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    • pp.307-317
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    • 2012
  • Objective: The purpose of this study was to investigate the isolation and characterization of multipotent human periodontal ligament (PDL) stem cells and to assess their ability to differentiate into bone, cartilage, and adipose tissue. Methods: PDL stem cells were isolated from 7 extracted human premolar teeth. Human PDL cells were expanded in culture, stained using anti-CD29, -CD34, -CD44, and -STRO-1 antibodies, and sorted by fluorescent activated cell sorting (FACS). Gingival fibroblasts (GFs) served as a positive control. PDL stem cells and GFs were cultured using standard conditions conducive for osteogenic, chondrogenic, or adipogenic differentiation. Results: An average of $152.8{\pm}27.6$ colony-forming units was present at day 7 in cultures of PDL stem cells. At day 4, PDL stem cells exhibited a significant increase in proliferation (p < 0.05), reaching nearly double the proliferation rate of GFs. About $5.6{\pm}4.5%$ of cells in human PDL tissues were strongly STRO-1-positive. In osteogenic cultures, calcium nodules were observed by day 21 in PDL stem cells, which showed more intense calcium staining than GF cultures. In adipogenic cultures, both cell populations showed positive Oil Red O staining by day 21. Additionally, in chondrogenic cultures, PDL stem cells expressed collagen type II by day 21. Conclusions: The PDL contains multipotent stem cells that have the potential to differentiate into osteoblasts, chondrocytes, and adipocytes. This adult PDL stem cell population can be utilized as potential sources of PDL in tissue engineering applications.

Changes and characteristics of the biochemical components on the differentiation of soybean cell tissue cultures : (II) Changes and characteristics of the lipid on the differentiation of soybean cell tissue cultures (대두 기내 배양체의 분화에 대한 생화학적 성분의 변화와 특성 : (II) 대두 기내 배양체의 분화에 대한 지질의 변화와 특성)

  • Nam, Sang-Hae;Yang, Min-Suk
    • Applied Biological Chemistry
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    • v.34 no.2
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    • pp.142-148
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    • 1991
  • When cultured soybean immature seed on 15th days after flowering, suitable temperature in formation of callus were $24{\sim}27^{\circ}C$, and embryogenic callus(EC) were generated at medium containing NAA with growth regulators, and then, results were subcultured the EC, a plenty of shoots and roots were formed at medium supplemented BA 2mg/1 and IAA 2mg/1, respectively, however when used at medium the same time supplemented BA 2mg/1 and IAA 2mg/1, formation of cullus was energetic, and a symptom of organization was not showed , Total lipid contents include in each cultures were increased at low temperature of cultural conditions as much as possible, but glycolipid, phospholipid, free sterol contents were a little increased at $24{\sim}27^{\circ}C$, and free sterol content was increased at a case of embryogenic structure were generated. In fatty acid compositions in each cultures, the contents of unsaturated fatty acid were plenty in EC, and unsaturation rate was 0.837. Besides, in sterol compositions, cholesterol content was remarkably high in EC than that of other cultures.

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INFLUENCE OF CO-CULTURED FIBROBLASTS ON THE DIFFERENTIATION OF MOUSE CALVARIA-DERIVED UNDIFFERENTIATED MESENCHYMAL CELLS IN VITRO (복합 및 유격배양한 섬유모세포가 마우스 두개관 미분화간엽세포의 골세포 분화에 미치는 영향)

  • Hwang, Yu-Sun;Kim, Myung-Rae
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.28 no.2
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    • pp.114-125
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    • 2002
  • This study was designed to evaluate the influence of fibroblasts or connective tissue from mouse oral mucosa on differentiation of neonatal mouse calvaria-derived osteoblasts and mineralization of bone nodules. Primary cell cultures from mouse calvarial osteoblasts and 2-4 passaged fibroblasts from oral mucosa were co-cultured in monolayer cultures, devided into 6 experimental group according to cell density or cell confluency. Osteoblasts were also co-cultured with fibroblasts in $Transwell^{(R)}$ culture plate with different co-cultured period according to osteoblast differentiation. The alkaline phosphatase activity were measured in monolayer cultures and cultures using $Transwell^{(R)}$. The mineralized bone nodules were presented by Von Kossa staining and density of mineralized nodules was measured by image analysis. The connective tissues with or without osteoblast seeding were cultured and examined histologically by Von Kossa and Trichrome Goldner staining. The results were as follows; 1. Prolonged maturation of matrix and delayed mineralization of bone nodules were resulted in monolayer cultures. 2. Co-culture of fibroblast with osteoblast using $Transwell^{(R)}$ during osteoblast proliferation stage stimulated proliferation of osteoblasts and increased alkaline phosphatase activity and mineralization of bone nodules. 3. Co-culture of fibroblast with osteoblast using $Transwell^{(R)}$ during matrix mineralization stage decreased and delayed mineralization of bone nodules. 4. In vitro cultured connective tissue with osteoblast seeding resulted in proliferation of osteoblasts and matrix formation with mineralization.

Propagation and Attenuation of Japanese Encephalitis Virus in Tissue Culture Cells (조직배양세포에서의 일본뇌염virus 증식에 관한 연구)

  • Lee, Ho-Wang;Moon, Seok-Bae
    • The Journal of the Korean Society for Microbiology
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    • v.16 no.1
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    • pp.83-89
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    • 1981
  • Japanese encephalitis has been prevalent for long time in the Far East and many patients have been reported in both South East and Mid-West Asia recently. Recently, vaccine was used in prevention of this viral disease of man which was derived from formalin inactivated virus inoculated into mouse brain, but live attenuated active vaccine for human is not developed yet. Author inoculated Japanese encephalitis virus into several cell culture strains for development of live attenuated encephalitis virus strain and the results were as follows: 1. Japanese encephalitis virus was inactivated rapidly in cell free medium at $36^{\circ}C$ and totally inactivated by 72 hours. 2. In growth curve of Japanese encephalitis virus in HeLa cell cultures, maximal multiplication of the virus was occured at 4th day and virus multiplication was continued for at least 12 days. 3. After succeeding passage of the virus in HeLa cell cultures and human esophagus epithelial cell cultures, infectivity of virus for mice was disappeared from 2nd passage in HeLa cell cultures and 3rd passage in esophagus epithelial cell cultures. 4. In inoculation to monkey kidney epithelial cells and chick embryo cell cultures, infectivity of the virus for mice was continued after 10th passages.

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Anticonvulsant potential of callus cultures of Convolvulus microphyllus Sieb.

  • Ahmad, Sayeed;Zafar, Rasheed-Uz;Shahid, Mohd
    • Advances in Traditional Medicine
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    • v.7 no.1
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    • pp.46-50
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    • 2007
  • Callus cultures of Convolvulus microphyllus Sieb. was induced on Murashige and Skoog's medium supplemented with 2,4-dichloro phenoxy acetic acid, 6-benzyl adenine, indole acetic acid and kinetin (1 ppm each). Methanolic extracts of whole plant, leaf, stem and leaf and stem calli were tested for anticonvulsant activity against standard drug phenytoin using maximal electroshock model on mice. It was observed that the animals treated with methanolic extracts of stem callus, leaf callus and whole plant (200 mg/kg, oral) showed significant protection against tonic convulsions induced by transcorneal electroshock. Anticonvulsant activity of methanolic extract of stem callus was comparable to that of standard drug phenytoin.

Immunohistochemical diagnosis of infectious pancreatic necrosis (어류 전염성훼장괴사증의 면역조직화학적 진단)

  • Kim, Soon-Bok
    • Korean Journal of Veterinary Pathology
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    • v.3 no.1
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    • pp.1-5
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    • 1999
  • This experiment was carried out to establish the immunohistochemical diagnostic method for infectious pancreatic necrosis in the monolayers of CHSE-214 cell cultures and paraffin-embedded tissue sections from rainbow trout infected with infectious pancreatic necrosis virus(IPNV). Specific identification of IPNV antigens was often demonstrated in the pancreatic exocrine cells, and less in the intestinal mucous epithelia and the renal hemopoietic tissues by the use of monoclonal antibodies against capsid protein VP2. The specific reaction was seen as a distinct red cytoplasmic color, often as small granules of various sizes. The result showed that streptavidin alkaline phosphatase immunohistochemisry specifically identified IPNV antigens in both infected cell cultures and tissue sections.

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Enzyme-linked immunosorbent assay (ELISA) for the detection of RVS (Retrovirus of Salmonid) (ELISA법을 이용한 연어과 어류의 RVS 검출(Retrovirus of Salmonid) 검출)

  • Oh, Myung-Joo;Yoshimizu, Mamoru
    • Journal of fish pathology
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    • v.9 no.2
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    • pp.169-176
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    • 1996
  • An indirect double antibody enzyme-linked immunosorbent assay (ELISA) was developed for rapid detection of a new virus isolated from abnormally swimming salmonid fish, RVS (Retrovirus of salmonid). Results using brain tissue homogenates, and infected cell cultures are described. The sensitivity of the methods is $10^{2.6}$ $TCID_{50}/100{\mu}l$ of the examined cell culture fluid. The specificity was confirmed by the ELISA inhibition test and virological examinations. Viral antigen could be detected in artificially infected fish tissue homogenates. The assay will allow the diagnosis of RVS-infected fish within a day.

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High Yield Saponin Production by Mass Cultures of Ginseng Transformed Tissue I. Induction, Culture of Transformed Tissue and Selection of High-Saponin-Producing Clones in Ginseng (인삼 형질전환 조직의 다량배양에 의한 Saponin 고 생산 I. 인삼에서 형질전환 조직의 유도, 배양과 Saponin 고 생산능주 선발)

  • 이정석;고경민
    • KSBB Journal
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    • v.9 no.2
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    • pp.157-164
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    • 1994
  • Hairy root clones of Panax ginseng were established by selection of some hairy roots formed on the leaf, stem and root segments transformed with Agrobacterium rhizogenes strain $A_4$. The transformed roots grew well in MS medium under the dark condition. To confirm the transformation with Ri-T-DNA, dot blot hybridization and opine analysis were Performed. Among four hairy roots induced from different part of ginseng, the HB3 hairy roots were examined for selection of high-saponin-producing clones. Four clones isolated from HB3 hairy root cultures displayed various phenotypes characterized by growth and total saponin content. Maximum growth was obtained for cultures of HB3-10 clone and the content of total saponin was 0.55 wt%. However, higher amount of total saponin was obtained with HB3-2 clone cultures(0.74 wt%) in spite of lower growth. Dot blot hybridization confirmed the introduction of Ri-T-DNA in the plant genome. In the opine test, agropine and mannopine were detected from all hairy root clones.

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