• Title/Summary/Keyword: culturing

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Growth Rate and Antimicrobial Activity of Lactobacillus plantarum Reacting with Bacterial Culture Supernatant (세균 배양액과 반응한 Lactobacillus plantarum의 생장속도 및 항균 활성에 대한 연구)

  • Ha, Eun-Mi
    • Microbiology and Biotechnology Letters
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    • v.43 no.3
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    • pp.187-194
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    • 2015
  • In this study, L. plantarum, when reacting with the culture media of potential pathogenic bacteria, exhibited an increase in growth rate and antimicrobial activity. In order to examine the characteristics and the nature of the reaction with the bacteria, this study carried out experiments involving culturing the test bacteria in M9 minimal media. Subsequently, the supernatant was incrassated by the decompression-drying method. Through colony forming unit assay, it was confirmed that L. plantarum had the function of growth inhibition to various bacteria. After culturing L. plantarum with bacterial media, the growth rate of L. plantarum was measured by absorbance (OD600), the results showed that the growth rate (E. coli treatment group: OD600 = 0.848, S. typhimurium treatment group: OD600 = 0.848) increased, as compared with the non-treated control group (OD600 = 0.48). In contrast, the concentrate itself did not induce the growth of L. plantarum. These results were observed as a universal phenomenon of the Lactobacillus species. Moreover, the increase in antimicrobial activity was observed in L. plantarum, which reacted with the culture media of E. coli and S. typhimurium, through a disc diffusion assay, and the result of growth inhibition against various bacteria was induced. Finally, based on the analysis results of the characteristics of bacteria culture media, which increased the growth rate of L. plantarum and antibacterial activity, the bacterial media had a tolerance for catabolic enzymes, pH 2−8 and heat. Therefore, this substance can be said to be a small molecule which is highly stable under various conditions.

Optimal Condition and Interspecific Cross-Reaction of H-Y Antibody Activity (H-Y항체활성의 최적조건과 종간교차반응)

  • ;H.S.Shim;J.B.Kim;H.Y.Park;K.S.Chung
    • Korean Journal of Animal Reproduction
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    • v.10 no.2
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    • pp.168-174
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    • 1986
  • These experiments were carried out to clarify the optimalconditions and interspecific cross reaction of H-Y antibody activity. H-Y antiserum was prepared in inbred SD female rats and Balb/c female mice by repeated immunization of rat newborn testis homogemate, rat and mouse spleen cells obtained from males of same strain. The activity of H-Y antibody in antiserum was tested by ELISA and biological tests. The cross reactivity of H-Y antibody was confirmed by culturing mouse and rabbit embryos in medium containing H-Y antibody and complement obtained from rat and guinea pig, respectively. The optimal condition for the activity of H-Y antibody was also investigated by culturing embryos in medium with different pH and complement concentration. The results obtained in these experiments were summarized as follows: 1. The formation rates of H-Y antibody in rats immunized with newborn testis and spleen cell were 40.0 and 50.0% respectively, and that in mouse immunized with spleen cell was 48.4%. 2. The activity of H-Y antibody was not affected by pH in range of 6.5 to 8.0, and the same was true for the relative concentration of complement to the H-Y antibody. 3. Minimum time needed for the activity of H-Y antibody was confirmed to be 0.5 to 1 hour and 24 to 48 hours respectively for the zona free embryos and intact embryos. 4. When mouse and rabbit embryos were treated with H-Y antibody obtained from rat, 46.4 and 54.8% of embryos were retarded or destroyed. From these results it could be said that H-Y antibody had strong interspecific cross reactivity.

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Application of a Dermal Equivalent to Organ Culture of Human Scalp Hair Follicle

  • Yoo, Bo-Young;Lee, Doo-Hoon;Seo, Young-Kwon;Shin, Youn-Ho;Song, Key-Yong;Seo, Seong-Jun;Whang, Sung-Joo;Kim, Young-Jin;Yang, Eun-Kyung;Park, Chang-Seo;Chang, Ih-Seop;Park, Jung-Keug
    • 한국생물공학회:학술대회논문집
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    • 2003.10a
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    • pp.359-362
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    • 2003
  • The recent development of methods for culturing hair follicles in vitro has proved an important tool to investigate many aspects of drug screening. Human hair follicle is composed of multiple types of cells, whose interactions regulate morphology and cycling-anagen, catagen, and telogen. Many investigators have tried to develop models to prolong of the period of hair elongation in vitro. However these are limited in submerged culture, which don't work due to the lack of cell-cell interactions which are abundant in vivo environment. So we applied dermal equivalent (DE) to culturing flair follicles to prolong hair growth period.

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Molecular Characterization of Neurally Differentiated Human Bone Marrow-derived Clonal Mesenchymal Stem Cells

  • Yi, TacGhee;Lee, Hyun-Joo;Cho, Yun-Kyoung;Jeon, Myung-Shin;Song, Sun U.
    • IMMUNE NETWORK
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    • v.14 no.1
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    • pp.54-65
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    • 2014
  • Bone marrow-derived mesenchymal stem cells (MSCs) are multipotent, with the ability to differentiate into different cell types. Additionally, the immunomodulatory activity of MSCs can downregulate inflammatory responses. The use of MSCs to repair injured tissues and treat inflammation, including in neuroimmune diseases, has been extensively explored. Although MSCs have emerged as a promising resource for the treatment of neuroimmune diseases, attempts to define the molecular properties of MSCs have been limited by the heterogeneity of MSC populations. We recently developed a new method, the subfractionation culturing method, to isolate homogeneous human clonal MSCs (hcMSCs). The hcMSCs were able to differentiate into fat, cartilage, bone, neuroglia, and liver cell types. In this study, to better understand the properties of neurally differentiated MSCs, gene expression in highly homogeneous hcMSCs was analyzed. Neural differentiation of hcMSCs was induced for 14 days. Thereafter, RNA and genomic DNA was isolated and subjected to microarray analysis and DNA methylation array analysis, respectively. We correlated the transcriptome of hcMSCs during neural differentiation with the DNA methylation status. Here, we describe and discuss the gene expression profile of neurally differentiated hcMSCs. These findings will expand our understanding of the molecular properties of MSCs and contribute to the development of cell therapy for neuroimmune diseases.

Radioprotective effects of dimethyl sulfoxide in the artificial skin reconstructed with cultured human cells (배양된 사람 세포를 이용해 제작한 인공피부에서 방사선조사에 대한 dimethyl sulfoxide의 보호 효과)

  • Ryu Young-Ha;Choi Karp-Shik;Song In-Hwan
    • Imaging Science in Dentistry
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    • v.32 no.1
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    • pp.11-18
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    • 2002
  • Purpose: To evaluate cultured human artificial skin as an experimental model for studying radiation effects in vitro. Materials and Methods: The skin was constructed by culturing keratinocytes over collagen lattice which made by culturing fibroblasts. Two groups were irradiated to gamma rays at single dose of 25 Gy with or without 3.5% of DMSO. Ultrastructures were investigated by electron microscopy after irradiation. The number of epidermal layers and expression of cytokeratin (CK) 14 & 10 were also seem by light microscopy. Results: At 2 days after irradiation in experimental group without DMSO, necrotic cells were rarely found in the spinosal layer and undercornified cells were visible in the homey layer. Similar findings were also found in experimental group with DMSO but in mild form. The number of epidermal layers in experimental group without DMSO were significantly fewer than other group. CK 14 expressed in all the layer excluding homey layer but CK 10 expressed over 3∼4 basal layers. Such patterns of CK expression were similar to all groups. It is suggested that structures of the keratinocytes and epidermal formation could be disturbed by irradiation in artificial skin and that DMSO can protect these damages. Conclusion : Therefore this work could be used as an organotypic experimental model in vitro using human cells for studying radiation effect in skin. Furthermore structural findings provided in this study could be used as useful basic data in further study using this model.

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A Comparison of Lyophilized Amniotic Membrane with Cryopreserved Amniotic Membrane for the Reconstruction of Rabbit Corneal Epithelium

  • Ahn Jae-Il;Jang In-Keun;Lee Doo-Hoon;Seo Young-Kwon;Yoon Hee-Hoon;Shin Youn-Ho;Kim Jae-Chan;Song Kye-Yong;Lee Hee-Gu;Yang Eun-Kyung;Kim Ki-Ho;Park Jung-Keung
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.3
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    • pp.262-269
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    • 2005
  • Many researchers have employed cryopreserved amniotic membrane (CAM) in the treatment of a severely damaged cornea, using corneal epithelial cells cultured on an amniotic membrane (AM). In this study, two Teflon rings were made for culturing the cells on the LAM and CAM, and were then used to support the AM, which is referred to in this paper as an Ahn's AM supporter. The primary corneal epithelial cells were obtained from the limbus, using an ex-plantation method. The corneal epithelium could be reconstructed by culturing the third­passage corneal epithelial cells on the AM. A lyophilized amniotic membrane (LAM) has a higher rate of graft take, a longer shelf life, is easier to store, and safer, due to gamma irradiation, than a (AM. The corneal epithelium reconstructed on the LAM and (AM, supported by the two­Teflon rings, was similar to normal corneal epithelium. However, the advantages of the LAM over that of the (AM make the former more useful. The reconstruction model of the corneal epithelium, using AM, is considered as a good in vitro model for transplantation of cornel epithelium into patients with a severely damaged cornea.

Rapid Detection of Mycobacterium tuberculosis Complex in Tissues by Using the Nested PCR (Nested PCR을 이용하여 조직으로부터 Mycobacterium tuberculosis Complex 신속검출)

  • Park, Jung-Yeon;Yang, Byoung-Seon
    • Korean Journal of Clinical Laboratory Science
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    • v.47 no.4
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    • pp.313-317
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    • 2015
  • Due to the increase in incidence of infection of Mycobacterium tuberculosis complex (MTC), it is imperative that a rapid diagnosis accompanies the handling of MTC. This is due to the three to eight weeks it takes to culture Mycobacteria, and the lack of sensitivity of microscopic examination of AFB. Recently, nested PCR has been used to detect and diagnose mycobacteria. It is especially useful in complementing diagnosis by histological extra pulmonary. After culturing all the specimens and practicing the nested PCR, we did comparison analysis between nested PCR and culture. There were 76 specimens, 31 of which were positive. Of the 31 positive specimens in culturing, only 22 were positive in nested PCR. Of the 45 negative specimens, 36 were negative in nested PCR. As a result, Sensitivity was 71% and specificity was 80%. Furthermore, the positive predictive value was 71% and negative predictive value was 80%. These results indicate that nested PCR based techniques are sensitive, specific, and rapid methods for the detection of MTC.

Pasteurella anatipestifer infection in ducklings in Korea (오리의 Pasteurella anatipestifer 감염증 발생)

  • Choi, Chung-ok;Kim, Gyoung-nyoun
    • Korean Journal of Veterinary Research
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    • v.33 no.1
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    • pp.93-99
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    • 1993
  • This study was conducted to investigate the cause of a new duck disease occured in southern part of Korea. A meat type duck farm located in Kangjin, Chonnam Province had experienced outbreaks of septicemic disease at around 20 days of age in nearly every batch of ducklings from early spring to early summer in 1989. Main symptoms of the birds were eye and nasal discharge, depression, inappetence, diarrhea and nervous signs such as tremor and ataxia. Some birds died suddenly without any signs. Mortality reached from 20% to 80% in severe cases. The autopsy findings of the affected ducklings revealed consistantly severe airsacculitis, fibropericarditis, perihepatitis and occasionaly enteritis and distended ureter with urate deposit. A rod shaped gram-negative bacterium was isolated purely from brain and liver of the diseased ducks by culturing the specimens on blood agar for 48 hours in candle jar. The isolate neither produced hemolysis nor grew on MacConKey Agar. It formed colony relatively slowly being recognizable at least 36 hours after culturing, reaching colony size of about 1mm in diameter at 48 hours culture. The colony looked iridescent under oblique light and had muddy odor. The isolate did not ferment carbohydrates tested but produced gelatinase, hippuricase and oxidase which were considered as characteristics of P anatipestifer. The isolate induced similar signs and lesions when infected experimentally into ducks of 3 to 38 days age via intraperitoneum or intratrachea. However it did not produce any clinical signs wen inoculated via intranasal route. It produced only mild signs in chicken just injected with a very large dose. The bacteria did not produce any signas or lesions in mice. It was concluded through biochemical and physiological tests and animal inoculation tests that the new disease was caused by P anatipestifer.

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Biosynthesis and Metabolism of Vitamin C in Suspension Cultures of Scutellaria baicalensis

  • Ahn, Young-Ock;Kwon, Suk-Yoon;Lee, Haeng-Soon;Park, Il-Hyun;Kwak, Sang-Soo
    • BMB Reports
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    • v.32 no.5
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    • pp.451-455
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    • 1999
  • The concentrations of L-ascorbic acid (AsA, ascorbate, vitamin C) and its biosynthetic and metabolically-related enzymes such as L-galactono-1,4-lactone dehydrogenase (GLDase), ascorbate peroxidase (APX), and ascorbate oxidase (ASO) were investigated in suspension cultures of Scutellaria baicalensis. Cells growing from 4 days after subculture (DAS) to 9 DAS and from 16 DAS to 19 DAS showed a diauxic growth, and then growth rapidly decreased with further culturing. The AsA content slowly increased to 19 DAS, reached a maximum at 21 DAS (ca $120\;{\mu}g/g$ dry cell wt), and then rapidly decreased with further culturing. GLDase and ASO activity were well correlated with the cell growth curve, showing a maximum at 19 DAS, whereas APX activity showed a good correlation with the changes in AsA content, showing a maximum at 21 DAS. The total ascorbate contents (reduced form, AsA, and oxidized form, dehydroascorbate, DHA) were markedly enhanced at 10 DAS when L-galactose and L-galactono-1,4-lactone (25 mM) were added to SH medium supplemented with 20 g/l sucrose at 9 DAS, by 5.5 and 6.8 times, respectively. DHA composed more than 90% of the total ascorbate contents in suspension cultures of S. baicalensis, even though the ratio of reduced to oxidized form slightly varied with cell growth stage. The results indicate that L-galactose and L-galactono-1,4-lactone are effective precursors of AsA in cell cultures of S. baicalensis, and that in vitro cultured cells provide suitable biomaterials for the study of biosynthesis and metabolism of AsA.

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Growth and Nutritional Composition of Eustigmatophyceae Monodus subterraneus and Nannochloropsis oceanica in Autotrophic and Mixotrophic Culture

  • Jo, Min Jin;Hur, Sung Bum
    • Ocean and Polar Research
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    • v.37 no.1
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    • pp.61-71
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    • 2015
  • Eicosapentaenoic acid (EPA) produced from marine organisms is widely used in nutraceuticals. Monodus subterraneus and Nannochloropsis oceanica, which are representative freshwater and marine Eustigmatophyceae, respectively, are known to have a high content of protein and lipid, particularly, EPA. In this study, to compare the growth and nutritional composition of M. subterraneus and N. oceanica, they were cultured in autotrophic and mixotrophic conditions with JM and f/2 medium, respectively, at $25^{\circ}C$. In addition, $80{\mu}mol\;photons\;m^{-2}s^{-1}$ with 24-hour and 12-hour light was provided, with the addition of 2% glucose to the medium for the mixotrophic culture. With regard to growth, M. subterraneus showed 10 times higher biomass in a mixotrophic culture than in an autotrophic one. However, no significant difference was observed for N. oceanica between the two culture methods. With respect to nutritional composition, M. subterraneus cultured autotrophically had a higher protein and lipid content, particularly EPA, than that cultured mixotrophically, but no significant difference was found in the two cultures of N. oceanica. Furthermore, M. subterraneus cultured autotrophically with continuous light showed higher nutritional composition, particularly EPA, than N. oceanica. In conclusion, the mass culture of freshwater M. subterraneus is much easier and more economical than marine N. oceanica. In addition, production of EPA will be economically improved if mixotrophic culturing of M. subterraneus is first conducted to maximize the biomass, and then secondary autotrophic culturing is performed.