• Title/Summary/Keyword: bioreactor culture

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Enhancement of BDNF Production by Co-cultivation of Human Neuroblastoma and Fibroblast Cells

  • Hong, Jong-Soo;Oh, Se-Jong;Kim, Sun-Hee;Park, Kwon-Tae;Cho, Jin-Sang;Park, Kyung-You;Lee, Jin-Ha;Lee, Hyeon-Yong
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.3 no.2
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    • pp.51-54
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    • 1998
  • It has been proved that co-cultivation of human neroblastoma cells and human fibroblast cells can enhance nerve cell growth and the production of BDNF in perfusion cultivation. In batch co-cultivation, maximum cell density was increased up to 1.76${\times}$106 viable cells/mL from 9${\times}$105 viable cells/mL of only neuroblastoma cell culture. The growth of neuroblastoma cells was greatly improved by culturing both nerve and fibroblast cells in a perfusion process, maintaining 1.5${\times}$106 viable cells/mL, which was much higher than that form fed-batch cultivation. The nerve cell growth was greatly enhance in both fed-batch and perfusion cultivations while the growth of fibroblast cells was not. It strongly implies that the factors secreted from human fibrobast cells and/or the environments of co-culture system can enhance both cell growth and BDNF secretion. Specific BDNF production rate was not enhanced in co-cultures; however, the production period was increased as the cell growth was lengthened in the co-culture case. Competitive growth between nerve cells and fibroblast cells was not observed in all cases, showing no changes of fibroblast cell growth and only enhancement of the neuroblastoma cell growth and overall BDNF production. It was also found that the perfusion cultivation was the most appropriate process for cultivating two cell lines simultaneously in a bioreactor.

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Production of casein phosphopeptides using Streptococcus faecalis var. liquefaciens cell immobilization (Streptococcus faecalis var. liquefaciens 전세포 고정화법을 이용한 Casein Phosphopeptides 생산)

  • Lee, Ki-Sung;Shin, Jae-Yoon;Jang, Yi-Hyun;Kweon, Dae-Hyuk;Park, Ki-Moon;Jin, Yong-Su
    • KSBB Journal
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    • v.23 no.1
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    • pp.59-64
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    • 2008
  • Optimum conditions for production of casein phosphopeptides (CPP) from sodium casenate by immobilized cell culture of Streptococcus faecalis var. liquefaciens were investigated. Immobilized cells were made by mixing 60% sodium alginate solution with an equal volume of culture broth at the end of exponential phase and subsequently dropping the mixture into $CaCl_{2}$ solution. Optimum conditions for CPP production by the immobilized cells were the same as those ($50^{\circ}C$, pH 7.0, and 10% substrate concentration) by the crude enzyme solution from the supernatant of culture broth. Optimum loading volume of the immobilized cells into a batch reactor was 30% (w/v). Using a continuous reactor loaded by the immobilized cells under the identified optimal conditions, we were able to produce CPP continuously up to 30 days with a maximum CPP conversion efficiency of 20%.

Production of Glutathione by yeast and Process Monitoring (효모에 의한 글루타치온의 생산과 공정 모니터링)

  • 김춘광;이종일
    • KSBB Journal
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    • v.19 no.3
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    • pp.192-199
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    • 2004
  • In this work the production of glutathione (GSH) by yeast Saccharomyces cerevisiae and the monitoring of the process were studied. In shaking culture the production of GSH was high at initial pH value of 4 and at temperature of 30$^{\circ}C$. But when L-cysteine was added to the culture medium at the beginning of the cultivation, the productivity of GSH was low. In case 0,5% (v/v) of L-cysteine, glycine and glutamic acid were introduced to the culture medium in the exponential cell growth phase, high concentration of GSH (about 90 mg/L) was produced in the bioreactor. A fed-batch operation with stepwise glucose feeding strategy allowed to produce 102 mg/L of GSH. The cultivation processes were on-line monitored by a 2-dimensional fluorescence sensor. A few off-line data such as cell growth, cystein concentration, phosphate concentration and GSH productivity could be well correlated to the fluorescence intensity of some combinations of excitation and emission wavelengths.

High-Density Cultivation of Microalgae using Microencapsulation (Microencapsulation에 의한 미세조류의 고밀도 배양)

  • HAN Young-Ho;LEE Jung-Suck;KWAK Jung-Ki;LEE Eung-Ho;CHO Man-Gi
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.32 no.2
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    • pp.186-191
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    • 1999
  • The three speices of miroalgae (Chlorella vulgaris, Dunaliella salina and Porphyridium purpureum) were immobilized in Ca-alginate capsules as a basic study for development of economic cultivation process, and then were cultivated in an air-bubble column bioreactor. Under the batch culture of aerobic conditions, the thickness of the capsule membrane and $CO_2$ supply did not affect the growth of the immobilized microalga, Chlorella vulgaris. Cell concentration of immobilized microalgae in the capsule was higher than those of imobilized microalgae in beads and free cells. The cell concentration of microencapsulated Dunaliella salina was greater about 5 times than that of free cells. Based on these results, it is concluded that the application of microencapsulation technology to the culture of microalgae was an effective method for high-density cultivation.

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Optimization of Culture Conditions for the [+]-Eudesmin Production in Magnolia Sieboldii Cells (함박꽃나무의 현탁배양세포로부터 [+]-Eudesmin의 생산을 위한 최적화)

  • Hwang Sung Jin
    • KSBB Journal
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    • v.20 no.1 s.90
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    • pp.34-39
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    • 2005
  • In order to product the furofuranoid lignans, (+)-eudesmin which is one of the secondary products from Magnolia sieboldii. through cell suspension cultures; various culture media, initial sucrose concentration, elicitations, shaking speeds, and inoculum sizes. Among the culture media tested, MS medium had a pronounced effect on suspension cell growth and (+)-eudesmin contents. The maximum dry cell weight (DCW) of 3.71 g per flask was obtained at inoculum size of 0.5 g and in MS medium supplemented with $3\%$ sucrose plus 0.5 mg/L 2,4-D after 8 weeks. (+)-Eudesmin biosynthesis was stimulated with high initial sucrose concentration ,and the maximum (+)-eudesmin production of $3.2{\mu}g/g$ DCW was achieved at 200mg/L chitosan and $5\%$ initial medium sucrose. The optimal shaking speeds for dry biomass accumulation and (+)-eudesmin contents was 130 rpm. This work is considered to be helpful for large-scale bioprocessing of Magnolia sieboldii suspension cell cultures in bioreactor.

Enhancement of Geldanamycin Production by pH Shock in Batch Culture of Streptomyces hygroscopicus subsp. duamyceticus

  • Song, Jae-Yang;Kim, Yoon-Jung;Hong, Young-Soo;Chang, Yong-Keun
    • Journal of Microbiology and Biotechnology
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    • v.18 no.5
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    • pp.897-900
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    • 2008
  • Various sequences of pH change were applied in a batch bioreactor to investigate pH shock effects on geldanamycin production by Streptomyces hygroscopicus subsp. duamyceticus JCM4427. In the control culture where the pH was not controlled, the maximum geldanamycin concentration was 414 mg/l. With the pHS1 mode of pH shock, that is, an abrupt pH change from pH 6.5 to pH 5.0 and then being maintained at around pH 5.0 afterward, 768 mg/l of geldanamycin was produced. With pHS2, in which the pH was changed sequentially from pH 6.7 to pH 5.0 and then back to pH 6.0, 429 mg/l of geldanamycin was produced. With pHS3 having a sequential pH change from pH 6.0 to pH 4.0 and then back to pH 6.5 followed by the third pH shock to pH 5.5, no geldanamycin production was observed. Considering that the productivity with pHS1 was about two-fold of that of the control culture with no pH control, we concluded that a more sophisticated manipulation of pH would further promote geldanamycin production.

Micropropagation of Lillium Oriental Hybrid 'Casa Blanca' using Bulblet Sections with swollen Basal Plate in Bioreactor (생물반응기에서 저반부가 비대된 자구 절편체에 의한 오리엔탈 나리 'Casa Blanca' 의 대량증식)

  • 한봉희;예병우;구대희
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.3
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    • pp.135-140
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    • 2001
  • A series of studies were carried out to establish micropropagation system, using airlift bioreactors (ebb $\varepsilon$ flood type, 5 L), of Lilium oriental hybrid 'Casa Blanca'. The bulblets with swollen basal plate were formed from bulb scales, then proliferated to bulblet clusters with swollen basal plate. Finally normal bulblets were formed from the sections. Bulblet formation and proliferation with swollen basal plate were not accomplished entirely in liquid culture of 5 L airlift bioreactors, but leafy bulb scales grew vigorously. Bulblet clusters with swollen basal plate were proliferated by periodic immersion culture. Bulblet proliferation was not affected by light, but scale leaves grew under light. MS medium containing 2.0 mg/L benzyl adenine (BA) and 0.3 mg/L indole acetic acid (IAA) was favorable to the bulblet proliferation with swollen basal plate. In liquid culture of 5 L bioreactors, bulblets from bulblet sections with swollen basal plate grew vigorously on MS medium with 70 g/L sucrose. It was effective for bulblet growth to replace the new medium after 8 weeks in culture during 16 weeks of cultural period. 15 g injection of bulblet sections as a cultural material was suitable for bulblet growth in 5 L bioreactors.

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Effects of Plant Growth Regulators, Medium Salt Strength and Nitrogen Ratio on Cell Culture of Gymmema sylvestre (식물생장조절물질, 무기물 농도 및 질소원 비율이 Gymmma sylvestre 세포 배양에 미치는 영향)

  • Lee, Eun-Jung;Han, Eun-Joo;Paek, Kee-Yoeup
    • Journal of Plant Biotechnology
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    • v.33 no.2
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    • pp.105-110
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    • 2006
  • This study was carried out to investigate the effects of plant growth regulators, medium salt strength and nitrogen ratio on cell culture of Gymnema sylvestre. Cell growth was inhibited by 2,4-D higher than 1.0 mg L$^{-1}$, but not by kinetin lower than 0.5 mg L$^{-1}$. Maximal cell growth was obtained at 1.0 mg L$^{-1}$ 2,4-D and 0.1 mg L$^{-1}$ kinetin. Cell growth was greatest at 1x MS medium but high strength of MS medium inhibited cell growth due to low water potential in the medium. In $NH_4^+:NO_3^-$ ratio of 0:60 (i.e. 0.0 mM $^NH_4^+$ and 60.0 mM $NO_3^-$), cells growth was highest but cells were smaller and whiter compared with those in other $NH_4^+:NO_3^-$ ratio. Reduced cell growth was observed with continuous culture. These results suggested that optimal cell culture of G. sylvestre could be achieved with 1x MS medium with 20:40 ratio of $NH_4^+:NO_3^-$ supplemented with 1.0 mg L$^{-1}$ 2,4-D and 0.1 mg L$^{-1}$ kinetin.

Mass Proliferation of Hibiscus hamabo Adventitious Root in an Air-lift Bioreactor, and the Antioxidant and Whitening Activity of the Extract (생물반응기를 이용한 황근 부정근의 대량증식과 추출물의 항산화 및 미백 활성 평가)

  • Lee, Jong-Du;Hyun, Ho Bong;Hyeon, Hyejin;Jang, Eunbi;Ko, Min-Hee;Yoon, Weon-Jong;Ham, Young Min;Jung, Yong-Hwan;Choi, Hwon;O, Eu Gene;Oh, Daeju
    • Korean Journal of Plant Resources
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    • v.35 no.4
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    • pp.435-444
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    • 2022
  • Hibiscus hamabo Sieb. et Zucc. (yellow hibiscus) is a deciduous semi-shrub plant and mainly growing in Jeju Island. This is known the unique wild hibiscus genus and classified as an 2nd grade of endangered plant for Korean Red List. In previous studies, properties of germination, ecological, genetical and salt resistance have been reported. In this study, we investigated mass-proliferated adventitious root using bioreactor, antioxidant and whitening effects to conduct functional ingredients. Yellow hibiscus were collected from Gujwa, Jeju by prior permission and they were introduced by explant type and various medium composition after surface sterilization. As a result, seed response rates were evaluated at range of 51.17~51.83%, in terms of comprehensive efficiency of shoot and root formation. In the case of adventitious root propagation condition was confirmed in half strength Murashige and Skoog medium salts, 30 mg/L sucrose, and 2 mg/L indole-3-butyric acid for 8 weeks in 5,000 mL bioreactor. We also compared between relationship with biomass and secondary metabolites accumulation by total phenolics content, the flavonoid content, DPPH free radical scavenging activity and melanin content. The results indicated that adventitious root mass proliferation, antioxidant and whitening effect could develop value of the high-quality cosmeceutical ingredient and further metabolite studies.

In vitro functional assenssment of bioartificial liver system using immobilized porcine hepatocyte spheroids

  • Lee, Ji-Hyun;Lee, Doo-Hoon;Yoon, Hee-Hoon;Jung, Doo-Hee;Park, Jung-Keug;Kim, Sung-Koo;Lee, Kwang-Woong;Lee, Suk-Koo
    • 한국생물공학회:학술대회논문집
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    • 2003.10a
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    • pp.305-306
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    • 2003
  • To treat fulminant hepatic failure (FHF) patients, various extracorporeal bioartificial liver (BAL) systems have been developed. Several requirements should be met for the development of BAL systems: hepatocytes should be cultured in a sufficiently high density; their metabolic functions should be of a sufficiently high level and duration; and the BAL systems module should permit scaling-up and aseptic handling. Several investigators have found that freshly isolated primary hepatocytes can be cultured into three dimensional, tightly packed, freely suspended, multicellular aggregates, or spheroids. These specialized cell structures exhibited enhanced liver specific functions and a prolonged differentiated state compared to cells maintained in a monolayer culture. Cells in spheroids appear to mimic the morphology and ultrastructure of the in vivo liver lobule. The ability of hepatocytes to organize into three-dimensional structures was hypothesized to contribute to their enhanced liver-specific activities. In this study, the ammonia removal rate and urea secretion rate of pig hepatocytes spheroids encapsulated in Ca-alginate bead were determined. A packed-bed bioreactor with encapsulated pig hepatocytes was devised as BAL support system. The efficacy of the system was evaluated in vitro.

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