The Experts below are selected from a list of 132 Experts worldwide ranked by ideXlab platform
Udo Reichl - One of the best experts on this subject based on the ideXlab platform.
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establishment of a mink enteritis vaccine production process in stirred tank reactor and wave bioreactor microcarrier culture in 1 10 l scale
Vaccine, 2007Co-Authors: Boris Hundt, N Schlawin, Holger Kassner, Yvonne Genzel, C. Best, Udo ReichlAbstract:Abstract A scale-up and process optimization scheme for the growth of adherent embryonic feline lung fibroblasts (E-FL) on microcarriers and the propagation of a mink enteritis virus (MEV) strain for the production of an inactivated vaccine is shown. Stirred-tank cultivations are compared with results obtained from Wave® Bioreactors. Transfer from a Roller Bottle-based production process into large-scale microcarrier culture with starting concentrations of 2 g/L Cytodex™ 1 microcarriers and 2.0 × 105 cells/mL was successful. A maximum cell yield of 1.2 × 106 cells/mL was obtained in stirred-tank microcarrier batch culture while cell numbers in the Wave® Bioreactor could not be determined accurately due to the fast sedimentation of microcarriers under non-rocking conditions required for sampling. Detailed off-line analysis was carried out to understand the behaviour of the virus–host cell system in both cultivation systems. Metabolic profiles for glucose, lactate, glutamine, and ammonium showed slight differences for both systems. E-FL cell growth was on the same level in stirred-tank and Wave® Bioreactor with a higher volumetric cell yield compared to Roller Bottles. Propagation of MEV, which can only replicate efficiently in mitotic cells, was characterized in the Wave® Bioreactor using a multiple harvest strategy. Maximum virus titres of 106.6 to 106.8 TCID50/mL were obtained, which corresponds to an increase in virus yield by a factor of about 10 compared to cultivations in Roller Bottles. As a consequence, a single Wave® Bioreactor cultivation of appropriate scale can replace hundreds of Roller Bottles. Thus, the Wave® Bioreactor proved to be a suitable system for large-scale production of an inactivated MEV vaccine.
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wave microcarrier cultivation of mdck cells for influenza virus production in serum containing and serum free media
Vaccine, 2006Co-Authors: Yvonne Genzel, Udo Reichl, R M Olmer, B SchaferAbstract:A process for equine influenza virus vaccine production using a microcarrier system (Cytodex 1) in a 2 L Wave bioreactor is described. Growth of Madin Darby canine kidney (MDCK) cells in serum containing GMEM medium (SC) is compared to growth in serum-free Ex-Cell MDCK medium (SF) without washing steps and medium exchange before infection. Cultivations with microcarrier concentrations of 2 and 4 g/L for both media are shown. Metabolic data from carbon and amino acid metabolism are discussed. Additionally, in Roller Bottle experiments the influence of multiplicity of infection (moi) and trypsin concentration on the HA value was investigated. Analysis of HA and TCID(50) at 37 degrees C showed a stable HA of maximum 2.6 log HA/100 microL for 2 weeks. Peak TCID(50) titers of 10(7.7) viruses/mL were achieved 20h post infection, but infectivity was below detection limit after 150 h. Cell attachment onto microcarriers under serum-free conditions was improved by Ca(2+) addition and by cell harvesting without trypsin using only an EDTA/PBS solution. For the wave cultivation maximum virus titers of 2.3-2.6 log HA units/100 microL were reached from infection with a moi of 0.05. However, in SF medium pH dropped to less than pH 6.8 which resulted in lower HA titers of 1.7 log HA units/100 microL. For the higher microcarrier concentration (4 g/L) medium exchange steps (500 mL) were needed for both media. Omission of the washing step and medium exchange before infection in SF medium clearly simplified the influenza production process; however, for higher virus yields a better pH control of the wave bioreactor would be required. Higher cell densities (2.8 x 10(6) cells/mL for 2 g/L microcarrier) and better attachment compared to stirred tank bioreactors showed, that the wave bioreactor is a good alternative to stirred tank processes for expanding production capacities in case of a pandemic.
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wave microcarrier cultivation of mdck cells for influenza virus production in serum containing and serum free media
Vaccine, 2006Co-Authors: Yvonne Genzel, Udo Reichl, R M Olmer, B SchaferAbstract:Abstract A process for equine influenza virus vaccine production using a microcarrier system (Cytodex 1) in a 2 L Wave bioreactor is described. Growth of Madin Darby canine kidney (MDCK) cells in serum containing GMEM medium (SC) is compared to growth in serum-free Ex-Cell MDCK medium (SF) without washing steps and medium exchange before infection. Cultivations with microcarrier concentrations of 2 and 4 g/L for both media are shown. Metabolic data from carbon and amino acid metabolism are discussed. Additionally, in Roller Bottle experiments the influence of multiplicity of infection (moi) and trypsin concentration on the HA value was investigated. Analysis of HA and TCID 50 at 37 °C showed a stable HA of maximum 2.6 log HA/100 μL for 2 weeks. Peak TCID 50 titers of 10 7.7 viruses/mL were achieved 20 h post infection, but infectivity was below detection limit after 150 h. Cell attachment onto microcarriers under serum-free conditions was improved by Ca 2+ addition and by cell harvesting without trypsin using only an EDTA/PBS solution. For the wave cultivation maximum virus titers of 2.3–2.6 log HA units/100 μL were reached from infection with a moi of 0.05. However, in SF medium pH dropped to less than pH 6.8 which resulted in lower HA titers of 1.7 log HA units/100 μL. For the higher microcarrier concentration (4 g/L) medium exchange steps (500 mL) were needed for both media. Omission of the washing step and medium exchange before infection in SF medium clearly simplified the influenza production process; however, for higher virus yields a better pH control of the wave bioreactor would be required. Higher cell densities (2.8 × 10 6 cells/mL for 2 g/L microcarrier) and better attachment compared to stirred tank bioreactors showed, that the wave bioreactor is a good alternative to stirred tank processes for expanding production capacities in case of a pandemic.
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serum free influenza virus production avoiding washing steps and medium exchange in large scale microcarrier culture
Vaccine, 2006Co-Authors: Yvonne Genzel, Udo Reichl, Marlies FischerAbstract:Abstract A complete serum-free process without washing steps and medium exchange before infection for influenza A virus vaccine production (equine and human) is described for cultivation in Roller Bottles and in a 5-L stirred tank microcarrier system. Adherent Madin–Darby canine kidney cells (MDCK) were adapted from growth in serum containing GMEM medium to growth in serum-free Ex-Cell MDCK medium. Roller Bottle experiments showed that the medium exchange step, typically required for serum containing vaccine production processes, could be omitted without losses in virus titre and without limitations in glucose or glutamine supply in the cultivation medium. The serum-free medium could even be used glutamine-free as it contained pyruvate, resulting in very low levels of ammonia. Cell attachment onto microcarriers was critical. Therefore, microcarriers had to be preconditioned in medium. Also, trypsin concentration used for inoculum preparation had to be reduced. After these modifications 1.3 × 106 cells/mL were obtained after 97 h (2 g/L Cytodex 1) of cell growth. Maximum virus titres of 2.3–2.9 log HA units/100 μL were obtained from infections with a multiplicity of infection (moi) of 0.05–0.10 for human and equine influenza A virus. Metabolite and amino acid profiles as well as on-line data for the serum-free process are compared with the serum containing process. Omission of the medium exchange before infection clearly simplified the process and reduced sterility risks due to washing steps.
Yvonne Genzel - One of the best experts on this subject based on the ideXlab platform.
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establishment of a mink enteritis vaccine production process in stirred tank reactor and wave bioreactor microcarrier culture in 1 10 l scale
Vaccine, 2007Co-Authors: Boris Hundt, N Schlawin, Holger Kassner, Yvonne Genzel, C. Best, Udo ReichlAbstract:Abstract A scale-up and process optimization scheme for the growth of adherent embryonic feline lung fibroblasts (E-FL) on microcarriers and the propagation of a mink enteritis virus (MEV) strain for the production of an inactivated vaccine is shown. Stirred-tank cultivations are compared with results obtained from Wave® Bioreactors. Transfer from a Roller Bottle-based production process into large-scale microcarrier culture with starting concentrations of 2 g/L Cytodex™ 1 microcarriers and 2.0 × 105 cells/mL was successful. A maximum cell yield of 1.2 × 106 cells/mL was obtained in stirred-tank microcarrier batch culture while cell numbers in the Wave® Bioreactor could not be determined accurately due to the fast sedimentation of microcarriers under non-rocking conditions required for sampling. Detailed off-line analysis was carried out to understand the behaviour of the virus–host cell system in both cultivation systems. Metabolic profiles for glucose, lactate, glutamine, and ammonium showed slight differences for both systems. E-FL cell growth was on the same level in stirred-tank and Wave® Bioreactor with a higher volumetric cell yield compared to Roller Bottles. Propagation of MEV, which can only replicate efficiently in mitotic cells, was characterized in the Wave® Bioreactor using a multiple harvest strategy. Maximum virus titres of 106.6 to 106.8 TCID50/mL were obtained, which corresponds to an increase in virus yield by a factor of about 10 compared to cultivations in Roller Bottles. As a consequence, a single Wave® Bioreactor cultivation of appropriate scale can replace hundreds of Roller Bottles. Thus, the Wave® Bioreactor proved to be a suitable system for large-scale production of an inactivated MEV vaccine.
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wave microcarrier cultivation of mdck cells for influenza virus production in serum containing and serum free media
Vaccine, 2006Co-Authors: Yvonne Genzel, Udo Reichl, R M Olmer, B SchaferAbstract:A process for equine influenza virus vaccine production using a microcarrier system (Cytodex 1) in a 2 L Wave bioreactor is described. Growth of Madin Darby canine kidney (MDCK) cells in serum containing GMEM medium (SC) is compared to growth in serum-free Ex-Cell MDCK medium (SF) without washing steps and medium exchange before infection. Cultivations with microcarrier concentrations of 2 and 4 g/L for both media are shown. Metabolic data from carbon and amino acid metabolism are discussed. Additionally, in Roller Bottle experiments the influence of multiplicity of infection (moi) and trypsin concentration on the HA value was investigated. Analysis of HA and TCID(50) at 37 degrees C showed a stable HA of maximum 2.6 log HA/100 microL for 2 weeks. Peak TCID(50) titers of 10(7.7) viruses/mL were achieved 20h post infection, but infectivity was below detection limit after 150 h. Cell attachment onto microcarriers under serum-free conditions was improved by Ca(2+) addition and by cell harvesting without trypsin using only an EDTA/PBS solution. For the wave cultivation maximum virus titers of 2.3-2.6 log HA units/100 microL were reached from infection with a moi of 0.05. However, in SF medium pH dropped to less than pH 6.8 which resulted in lower HA titers of 1.7 log HA units/100 microL. For the higher microcarrier concentration (4 g/L) medium exchange steps (500 mL) were needed for both media. Omission of the washing step and medium exchange before infection in SF medium clearly simplified the influenza production process; however, for higher virus yields a better pH control of the wave bioreactor would be required. Higher cell densities (2.8 x 10(6) cells/mL for 2 g/L microcarrier) and better attachment compared to stirred tank bioreactors showed, that the wave bioreactor is a good alternative to stirred tank processes for expanding production capacities in case of a pandemic.
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wave microcarrier cultivation of mdck cells for influenza virus production in serum containing and serum free media
Vaccine, 2006Co-Authors: Yvonne Genzel, Udo Reichl, R M Olmer, B SchaferAbstract:Abstract A process for equine influenza virus vaccine production using a microcarrier system (Cytodex 1) in a 2 L Wave bioreactor is described. Growth of Madin Darby canine kidney (MDCK) cells in serum containing GMEM medium (SC) is compared to growth in serum-free Ex-Cell MDCK medium (SF) without washing steps and medium exchange before infection. Cultivations with microcarrier concentrations of 2 and 4 g/L for both media are shown. Metabolic data from carbon and amino acid metabolism are discussed. Additionally, in Roller Bottle experiments the influence of multiplicity of infection (moi) and trypsin concentration on the HA value was investigated. Analysis of HA and TCID 50 at 37 °C showed a stable HA of maximum 2.6 log HA/100 μL for 2 weeks. Peak TCID 50 titers of 10 7.7 viruses/mL were achieved 20 h post infection, but infectivity was below detection limit after 150 h. Cell attachment onto microcarriers under serum-free conditions was improved by Ca 2+ addition and by cell harvesting without trypsin using only an EDTA/PBS solution. For the wave cultivation maximum virus titers of 2.3–2.6 log HA units/100 μL were reached from infection with a moi of 0.05. However, in SF medium pH dropped to less than pH 6.8 which resulted in lower HA titers of 1.7 log HA units/100 μL. For the higher microcarrier concentration (4 g/L) medium exchange steps (500 mL) were needed for both media. Omission of the washing step and medium exchange before infection in SF medium clearly simplified the influenza production process; however, for higher virus yields a better pH control of the wave bioreactor would be required. Higher cell densities (2.8 × 10 6 cells/mL for 2 g/L microcarrier) and better attachment compared to stirred tank bioreactors showed, that the wave bioreactor is a good alternative to stirred tank processes for expanding production capacities in case of a pandemic.
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serum free influenza virus production avoiding washing steps and medium exchange in large scale microcarrier culture
Vaccine, 2006Co-Authors: Yvonne Genzel, Udo Reichl, Marlies FischerAbstract:Abstract A complete serum-free process without washing steps and medium exchange before infection for influenza A virus vaccine production (equine and human) is described for cultivation in Roller Bottles and in a 5-L stirred tank microcarrier system. Adherent Madin–Darby canine kidney cells (MDCK) were adapted from growth in serum containing GMEM medium to growth in serum-free Ex-Cell MDCK medium. Roller Bottle experiments showed that the medium exchange step, typically required for serum containing vaccine production processes, could be omitted without losses in virus titre and without limitations in glucose or glutamine supply in the cultivation medium. The serum-free medium could even be used glutamine-free as it contained pyruvate, resulting in very low levels of ammonia. Cell attachment onto microcarriers was critical. Therefore, microcarriers had to be preconditioned in medium. Also, trypsin concentration used for inoculum preparation had to be reduced. After these modifications 1.3 × 106 cells/mL were obtained after 97 h (2 g/L Cytodex 1) of cell growth. Maximum virus titres of 2.3–2.9 log HA units/100 μL were obtained from infections with a multiplicity of infection (moi) of 0.05–0.10 for human and equine influenza A virus. Metabolite and amino acid profiles as well as on-line data for the serum-free process are compared with the serum containing process. Omission of the medium exchange before infection clearly simplified the process and reduced sterility risks due to washing steps.
Paul Martin - One of the best experts on this subject based on the ideXlab platform.
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analysis of the tissue movements of embryonic wound healing dii studies in the limb bud stage mouse embryo
Developmental Biology, 1995Co-Authors: Jane Mccluskey, Paul MartinAbstract:Abstract The tissue movements of epithelial spreading and mesenchymal contraction play key roles in many aspects of embryonic morphogenesis. One way of studying these movements in a controlled manner is to make all excisional skin wound to an embryo and watch the wound heal. In this paper we report our studies of healing of a simple excisional lesion made to the limb bud stage mouse embryo. The wounded, living embryo is cultured in a Roller Bottle; under such conditions the wound heals with a highly reproducible time course and is completely closed by 24 hr. During the healing period the environment bathing the wound can be simply manipulated by adding drugs or factors to the culture medium. We have used DiI to label mesenchymal cells exposed at the margin of the initial wound and, by following their fate and measuring the area of mesenchyme remaining exposed at various time points during the healing process, we have quantified both the extent of mesenchymal contraction and the extent of reepithelialisation by movement of epidermis over mesenchyme. We show that the two types of tissue movement contribute almost equally (50:50) to the total wound closure rate. We have gone on to investigate the cell machinery underlying these processes. In adult wounds the epidermis migrates by means of lamellipodial crawling, but we show that reepithelialisation in the embryo is achieved instead by purse-string contraction of a cable of filamentous actin which assembles in the basal layer of cells at the free edge of the epidermis. Addition of cytochalasin D to the culture medium blocks formation of this actin cable end leads to failure of reepithelialisation. Contraction of adult wound connective tissue appears to be driven by conversion of dermal fibroblasts into a specialist smooth muscle-like fibroblast, the myofibroblast. However, using an antibody recognising the αisoform of smooth muscle actin and specific for smooth muscle cells and myofibroblasts, we show that a similar conversion into myofibroblasts does not occur at any stage during the embryonic wound healing process. These observations indicate that both of the tissue movements of embryonic wound healing utilise cell machinery fundamentally different from that driving the analogous tissue movements of adult healing.
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analysis of the tissue movements of embryonic wound healing dii studies in the limb bud stage mouse embryo
Developmental Biology, 1995Co-Authors: Jane T Mccluskey, Paul MartinAbstract:The tissue movements of epithelial spreading and mesenchymal contraction play key roles in many aspects of embryonic morphogenesis. One way of studying these movements in a controlled manner is to make an excisional skin wound to an embryo and watch the wound heal. In this paper we report our studies of healing of a simple excisional lesion made to the limb bud stage mouse embryo. The wounded, living embryo is cultured in a Roller Bottle; under such conditions the wound heals with a highly reproducible time course and is completely closed by 24 hr. During the healing period the environment bathing the wound can be simply manipulated by adding drugs or factors to the culture medium. We have used DiI to label mesenchymal cells exposed at the margin of the initial wound and, by following their fate and measuring the area of mesenchyme remaining exposed at various time points during the healing process, we have quantified both the extent of mesenchymal contraction and the extent of reepithelialisation by movement of epidermis over mesenchyme. We show that the two types of tissue movement contribute almost equally (50:50) to the total wound closure rate. We have gone on to investigate the cell machinery underlying these processes. In adult wounds the epidermis migrates by means of lamellipodial crawling, but we show that reepithelialisation in the embryo is achieved instead by purse-string contraction of a cable of filamentous actin which assembles in the basal layer of cells at the free edge of the epidermis. Addition of cytochalasin D to the culture medium blocks formation of this actin cable and leads to failure of reepithelialisation. Contraction of adult wound connective tissue appears to be driven by conversion of dermal fibroblasts into a specialist smooth muscle-like fibroblast, the myofibroblast. However, using an antibody recognising the alpha-isoform of smooth muscle actin and specific for smooth muscle cells and myofibroblasts, we show that a similar conversion into myofibroblasts does not occur at any stage during the embryonic wound healing process. These observations indicate that both of the tissue movements of embryonic wound healing utilise cell machinery fundamentally different from that driving the analogous tissue movements of adult healing.
R M Olmer - One of the best experts on this subject based on the ideXlab platform.
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wave microcarrier cultivation of mdck cells for influenza virus production in serum containing and serum free media
Vaccine, 2006Co-Authors: Yvonne Genzel, Udo Reichl, R M Olmer, B SchaferAbstract:A process for equine influenza virus vaccine production using a microcarrier system (Cytodex 1) in a 2 L Wave bioreactor is described. Growth of Madin Darby canine kidney (MDCK) cells in serum containing GMEM medium (SC) is compared to growth in serum-free Ex-Cell MDCK medium (SF) without washing steps and medium exchange before infection. Cultivations with microcarrier concentrations of 2 and 4 g/L for both media are shown. Metabolic data from carbon and amino acid metabolism are discussed. Additionally, in Roller Bottle experiments the influence of multiplicity of infection (moi) and trypsin concentration on the HA value was investigated. Analysis of HA and TCID(50) at 37 degrees C showed a stable HA of maximum 2.6 log HA/100 microL for 2 weeks. Peak TCID(50) titers of 10(7.7) viruses/mL were achieved 20h post infection, but infectivity was below detection limit after 150 h. Cell attachment onto microcarriers under serum-free conditions was improved by Ca(2+) addition and by cell harvesting without trypsin using only an EDTA/PBS solution. For the wave cultivation maximum virus titers of 2.3-2.6 log HA units/100 microL were reached from infection with a moi of 0.05. However, in SF medium pH dropped to less than pH 6.8 which resulted in lower HA titers of 1.7 log HA units/100 microL. For the higher microcarrier concentration (4 g/L) medium exchange steps (500 mL) were needed for both media. Omission of the washing step and medium exchange before infection in SF medium clearly simplified the influenza production process; however, for higher virus yields a better pH control of the wave bioreactor would be required. Higher cell densities (2.8 x 10(6) cells/mL for 2 g/L microcarrier) and better attachment compared to stirred tank bioreactors showed, that the wave bioreactor is a good alternative to stirred tank processes for expanding production capacities in case of a pandemic.
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wave microcarrier cultivation of mdck cells for influenza virus production in serum containing and serum free media
Vaccine, 2006Co-Authors: Yvonne Genzel, Udo Reichl, R M Olmer, B SchaferAbstract:Abstract A process for equine influenza virus vaccine production using a microcarrier system (Cytodex 1) in a 2 L Wave bioreactor is described. Growth of Madin Darby canine kidney (MDCK) cells in serum containing GMEM medium (SC) is compared to growth in serum-free Ex-Cell MDCK medium (SF) without washing steps and medium exchange before infection. Cultivations with microcarrier concentrations of 2 and 4 g/L for both media are shown. Metabolic data from carbon and amino acid metabolism are discussed. Additionally, in Roller Bottle experiments the influence of multiplicity of infection (moi) and trypsin concentration on the HA value was investigated. Analysis of HA and TCID 50 at 37 °C showed a stable HA of maximum 2.6 log HA/100 μL for 2 weeks. Peak TCID 50 titers of 10 7.7 viruses/mL were achieved 20 h post infection, but infectivity was below detection limit after 150 h. Cell attachment onto microcarriers under serum-free conditions was improved by Ca 2+ addition and by cell harvesting without trypsin using only an EDTA/PBS solution. For the wave cultivation maximum virus titers of 2.3–2.6 log HA units/100 μL were reached from infection with a moi of 0.05. However, in SF medium pH dropped to less than pH 6.8 which resulted in lower HA titers of 1.7 log HA units/100 μL. For the higher microcarrier concentration (4 g/L) medium exchange steps (500 mL) were needed for both media. Omission of the washing step and medium exchange before infection in SF medium clearly simplified the influenza production process; however, for higher virus yields a better pH control of the wave bioreactor would be required. Higher cell densities (2.8 × 10 6 cells/mL for 2 g/L microcarrier) and better attachment compared to stirred tank bioreactors showed, that the wave bioreactor is a good alternative to stirred tank processes for expanding production capacities in case of a pandemic.
Arildo Jose Braz De Oliveira - One of the best experts on this subject based on the ideXlab platform.
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adventitious root culture of pfaffia glomerata spreng pedersen in a Roller Bottle system an alternative source of β ecdysone
Phytochemistry Letters, 2021Co-Authors: Thaila Fernanda Oliveira Da Silva, Arildo Jose Braz De Oliveira, Cristina Sayuri Yamaguchi, Susana Tavares Cotrim Ribeiro, Alexandre Da Silva Avincola, Eduardo Jorge Pilau, Carla Porto, Regina Aparecida Correia GoncalvesAbstract:Abstract β-ecdysone is the main compound of commercial interest produced by the roots of Pfaffia glomerata (Spreng.) Pedersen, and is used in herbal medicines and dietary supplements. In-vitro root culture facilitates the continuous production of high-quality compounds, and uses sustainable production techniques. Adventitious roots of P. glomerata were cultivated in a Roller Bottle system and an orbital gyratory system, both in the absence of light. The quantification of β-ecdysone by Liquid Chromatography-Mass Spectrometry/Mass Spectrometry (LC–MS/MS) revealed that the highest accumulation of metabolites occurred after 8 weeks in the Roller Bottle culture system, wherein 1.81 μg of β-ecdysone was recovered from 30 mg of dry root. These results suggest that adventitious root culture in-vitro may be an alternative strategy for β-ecdysone production, and demonstrates the potential use of bioreactors for practical applications.
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chemical characterization and prebiotic activity of fructo oligosaccharides from stevia rebaudiana bertoni roots and in vitro adventitious root cultures
Carbohydrate Polymers, 2016Co-Authors: Sheila Mara Sanches Lopes, Regina Aparecida Correia Goncalves, Eduardo Jorge Pilau, Mariane Grigio Francisco, Bruna Higashi, Rafaela Takako Ribeiro De Almeida, Gabriela Krausova, Jose Eduardo Goncalves, Arildo Jose Braz De OliveiraAbstract:Stevia rebaudiana (Bertoni) is widely studied because of its foliar steviol glycosides. Fructan-type polysaccharides were recently isolated from its roots. Fructans are reserve carbohydrates that have important positive health effects and technological applications in the food industry. The objective of the present study was to isolate and characterize fructo-oligosaccharides (FOSs) from S. rebaudiana roots and in vitro adventitious root cultures and evaluate the potential prebiotic effect of these molecules. The in vitro adventitious root cultures were obtained using a Roller Bottle system. Chemical analyses (gas chromatography-mass spectrometry, (1)H nuclear magnetic resonance, and off-line electrospray ionization-mass spectrometry) revealed similar chemical properties of FOSs that were obtained from the different sources. The potential prebiotic effects of FOSs that were isolated from S. rebaudiana roots enhanced the growth of both bifidobacteria and lactobacilli, with strains specificity in their fermentation ability.
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establishment of adventitious root culture of stevia rebaudiana bertoni in a Roller Bottle system
Plant Cell Tissue and Organ Culture, 2011Co-Authors: Rafael Valim Reis, Ana Paula Patrao Luis Borges, Talita Perez Cantuaria Chierrito, Eliezer Rodrigues De Souto, Lauro Mera De Souza, Marcello Iacomini, Arildo Jose Braz De Oliveira, Regina Aparecida Correia GoncalvesAbstract:Cultures of adventitious roots of Stevia rebaudiana (Bert.) Bertoni were performed in a Roller Bottle system for the production of both primary and secondary metabolites. Adventitious roots were induced from 1-cm-long root tip explants derived from in vitro regenerated plantlets on solid Murashige and Skoog (MS 1962) media supplemented with 10.7 μM of α-naphthaleneacetic acid. These cultures were successfully maintained in the same medium for 6 months with regular subcultures after 4 weeks. Thereafter, the roots were cut into 1.0- to 1.5-cm-long segments and transferred to the Roller Bottle system containing a fresh root tissue culture on liquid MS medium supplemented with 10.7 μM NAA. The apparatus consisted of a flask rolling system adjusted to 4g, and 3° of flask inclination. The roots were allowed to grow in the absence of light for adaptation and adventitious root formation. The best conditions for cultivation were investigated, considering culture volume (25 ml), culture period (4 weeks), salt concentrations in the nutrient medium (33%) and optimal initial inoculum (0.2 g) of S. rebaudiana roots. These results could give important information on how to improve the development of adventitious roots of S. rebaudiana for the production of primary and secondary metabolites.
