• 제목/요약/키워드: Suspension Culture

검색결과 569건 처리시간 0.029초

Suspension culture of Stably Transformed Drosophila melanogaster S2 Cells expressing EGFP and EPO

  • Sohn, Bong-Hee;Lee, Jong-Min;Kim, Yong-Soon;Kang, Pil-Don;Lee, Sang-Uk;Chung, In-Sik
    • 한국잠사학회:학술대회논문집
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    • 한국잠사학회 2003년도 제46회 춘계 학술연구 발표회
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    • pp.40-40
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    • 2003
  • Recombinant plasmids harboring heterologous genes coding enhanced green fluorescent protein (EGFP) and erythropoietin (EPO) were transfected and expressed in Drosophila melanogaster S2 cells. Stably transformed cell populations expressing EGFP or monkey EPO were isolated after 4 weeks of selection with hygromycin B. (omitted)

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형질전환된 담배 세포 현탁배양을 이용한 hGM-CSF 생산에서 ultrasound가 미치는 영향

  • 명현종;이기용;김동일
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2003년도 생물공학의 동향(XII)
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    • pp.284-287
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    • 2003
  • 본 연구에서는 hGM-CSF를 생산하는 형질전환된 담배세포배양에서 ultrasound가 세포생장과 hGM-CSF 생산에 미치는 영향을 조사하였다. 세포의 생장은 ultrasound에 노출된 직후 약간의 저해를 보이지만 2일후에 정상의 세포와 비슷한 수준으로 회복되었다. 세포 외부로 분비되는 hGM-CSF는 ultrasound에 노출된 직후 증가하고 그 이후 감소하다 배양후반에 증가를 보였다. 세포내부에서 생산되는 hGM-CSF는 ultrasound 처리 이후 전반적으로 대조구보다 많이 생산됨을 보였다. 총 생산된 hGM-CSF는ultrasound에 노출 시킨 배양 6일째 $34.9\;{\mu}g/L$로 최대였으며 대조구보다 31.5%의 증가를 보였다.

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Large-Scale Culture of Hepatitis A Virus in Human Diploid MRC-5 Cells and Partial Purification of the Viral Antigen for Use as a Vaccine

  • Kim, Hyun-Seok;Chung, Yong-Ju;Jeon, Yeong-Joong;Lee, Sung-Hee
    • Journal of Microbiology and Biotechnology
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    • 제9권4호
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    • pp.386-392
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    • 1999
  • A large-scale culture of hepatitis A virus in human diploid MRC-5 cells was conducted. In a roller bottle culture, the virus was grown to a maximum titer in 3 weeks after infection. Over 95% of the cell-associated virus was excreted after culturing the infected cells in suspension media without fetal bovine serum for 3 days. The cultured virus was inactivated with formalin, concentrated by ultrafiltration, and partially purified by ultracentrifugation in a non-ionic gradient medium of Renocal. Two separate peak fractions showing high anti-HAY ELISA titer were pooled and about 40% of HAV antigen was recovered by this purification procedure. Of the partially purified vaccine, the protein pattern in SDS-PAGE and immunogenicity in mice were compared with a commercial HAV vaccine. In SDS-PAGE, the purified vaccine in this study and the commercial vaccine showed almost the same protein pattern. The seroconversion rate of the purified vaccine in mice was not different from that of the commercial vaccine. Therefore, we could prepare a good grade of HAV vaccine by a simple purification procedure although the purification itself was not completed.

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세포 현탁배양에 의한 현호색 알칼로이드의 생산 (Studies on Production of Alkaloid by Plant Cell Culture of Corydalis remota)

  • 장정인;지형준;신승원
    • 생약학회지
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    • 제27권4호
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    • pp.289-294
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    • 1996
  • In previous paper, we described the induced callus of Corydalis remota contains a significant amount of alkaloids. This study describes an optimal condition to maximize alkaloid production. The suspension cultures maintained alkaloid production ability after fifth subculture and a small amount of alkaloid seemed to be released out of cells. The yields of alkaloid by cultured cells was varied depending on the concentrations of NAA, carbon sources and phosphate ion and depending on the vitamin combinations and concentrations. Biosynthetic precursor and an elicitor treatment also affected the total alkaloid yield of the cultures. The optimal conditions for alkaloid production were as follows: 1) MS basal salt containing 30 g/l of glucose, 1.0 mg/l of NAA, and vitamins of LS medium should be used. 2) The culture should be treated with tyrosine 20 mg/l, and yeast extract 1.5 ml/l after the culture reached a stationary phase of growth. Five alkaloids were isolated from the cultures and they were characterized. The spectral data unambiguously revealed that the isolated compounds were dihydrosanguinarine, protopine. tetrahydropalmatine, allocyptopine and ambinine, respectively.

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Combined TGE-SGE Expression of Novel PAI-1-Resistant t-PA in CHO DG44 Cells Using Orbitally Shaking Disposable Bioreactors

  • Davami, Fatemeh;Barkhordari, Farzaneh;Alebouyeh, Mahmoud;Adeli, Ahmad;Mahboudi, Fereidoun
    • Journal of Microbiology and Biotechnology
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    • 제21권12호
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    • pp.1299-1305
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    • 2011
  • An important modification of thrombolytic agents is resistance to plasminogen activator inhibitor-1 (PAI-1). In previous studies, a new truncated PAI-1-resistant variant was developed based on deletion of the first three domains in t-PA and the substitution of KHRR 128-131 amino acids with AAAA in the truncated t-PA. The novel variant expressed in a static culture system of Chinese Hamster Ovary (CHO) DG44 cells exhibited a higher resistance to PAI-1 when compared with the full-length commercial drug; Actylase. In the present study, the truncated-mutant protein was expressed in CHO DG44 cells in 50 ml orbital shaking bioreactors. The final yield of the truncated-mutant in the culture was 752 IU/ml, representing a 63% increase compared with the static culture system. Therefore, these results suggest that using the combined features of a transient and stable expression system is feasible for the production of novel recombinant proteins in the quantities needed for preclinical studies.

Effect of Pluronic F-68 and Oxygen Vectors on Cell Growth of Angelica gigas Nakai in Fed-batch Culture

  • 전수환;이상윤;조지숙;민병혁;김동일
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2000년도 추계학술발표대회 및 bio-venture fair
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    • pp.159-162
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    • 2000
  • 유가식배양을 통한 고농도배양에서 계면활성제와 산소전달물질의 첨가는 세포의 고밀도와 배지의 고점도로 인해 발생하는 nutrient의 물질전달제한과 산소부족현상을 효과적으로 해결하였다. Pluronic 10R-5는 Pluronic F-68과 같은 triblock copolymer 구조이며 세포생장의 저해를 나타내지 않았다. 그러나 diblock copolymer구조이며 소수성 부분이 hydrocarbon block인 Plurafac A-38은 세포의 생장을 저해함을 관찰할 수 있었다. 따라서 장기간의 유가식배양이나 perfusion 배양을 수행할 경우 세포의 생존도를 증대시켜 유용물질의 생산성을 높일 수 있으며 여러 형태의 bioreactor에서 고농도 배양을 가능케 하여 유용물질 생산성을 높일수 있을 것이라고 기대된다.

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식물세포배양에 의한 Corydalis Alkaloid의 생산(I) (Production of Corydalis Alkaloids by Plant Cell Culture(I))

  • 장정인;신승원;지형준
    • 생약학회지
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    • 제26권4호
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    • pp.419-425
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    • 1995
  • Corydalis remota Fish. ex Max. (Papaveraceae) is a well known medicinal plant being used as analgesics or anticonvulsive in oriental medicine. As the alkaloid content is known to vary depending on the environmental factors, the technology of plant tissue culture can be adopted as source of Corydalis-alkaloids. The present study describes an establishment of tissue cultures of Corydalis which produce alkaloids consistently. Callus were induced from immature seeds of Corydalis remota by placing the seeds on MS static media containing NAA(0.25, 1.0 and 4.0 mg/l, respectively). The combined treatment of NAA(1.0 mg/l) with cytokinin(BAP 0.5 mg/l) improved the induction of callus. TLC scanning data followed by sequential extraction and purification revealed that the induced callus contains a significant amount of alkaloids. Cell suspension cultures were established by transferring the induced callus into the liquid media with the same condition of plant growth regulators as the callus culture.

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