• 제목/요약/키워드: Chinese Hamster Ovary

검색결과 212건 처리시간 0.026초

Zinc and Selenium Requirements for Glutathione Peroxidase Activity and Cell Survival in Chinese Hamster Ovary Cells Overexpressing Metallothionein

  • Kwun, In-Sook;John R. Arthur;John H. Beattie
    • Preventive Nutrition and Food Science
    • /
    • 제8권1호
    • /
    • pp.36-39
    • /
    • 2003
  • Many defined cell culture media were formulated over 3() years ago and may be deficient in certain micronutrients whose essentiality has only subsequently been recognised. The objective of this study was to evaluate whether alpha-minimal essential medium (MEM) supplemented with 10% foetal bovine serum contained sufficient selenium for optimal activity of the selenium containing enzymes cytosolic glutathione peroxidase (cGPx) and phospholipid hydroperoxide glutathione peroxidase (PHGPx) in cultured Chinese hamster ovary (CHO) cells. Additionally, the effect of zinc deficiency and metallothionein (MT) overexpression on cGPx and PHGPx activity was studied. The addition of 100 nM of selenous acid to the culture medium increased cGPx expression by 10-fold and PHGPx by about 2-fold in both wild-type CHO-K1 cells and CHO-K1 cells overexpressing mouse MT-1. Zinc deficiency had no significant effect on enzyme activity, but cells overexpressing mouse MT-1 had higher levels of cGPx activity. Zinc deficiency decreased cell survival but overexpression of MT-1 was partially protective, probably because its presence in quantity favoured the uptake, sequestration and cellular retention of any remaining zinc. This study demonstrates that selenium in complete alpha-MEM is insufficient for optimal cGPx and PHGPx activity and may compromise the cellular response to oxidative stress.

Changes in Cell Cycle-related Genes Expression of Chineses Hamster Ovary Cells cultured using Serum-free Media

  • Seo, Sung-Keum;Park, Hong-Woo;Choe, Tae-Boo
    • 한국생물공학회:학술대회논문집
    • /
    • 한국생물공학회 2003년도 생물공학의 동향(XIII)
    • /
    • pp.153-158
    • /
    • 2003
  • The genome-wide program of gene expression during the cell division cycle response to serum free media in chinese hamster ovary(CHO) cells was characterized using cDNA microarrays. Many transcripts of genes showed variation during the cell cycle. Characterization is critical step toward understanding the basic cell cycle processes and serum-free media role in CHO cells

  • PDF

Yeast hydrolysate as a low-cost additive to serum-free medium for the production of human thrombopoietin in suspension cultures of Chinese hamster ovary cells

  • 성윤희;이균민
    • 한국생물공학회:학술대회논문집
    • /
    • 한국생물공학회 2003년도 생물공학의 동향(XII)
    • /
    • pp.105-108
    • /
    • 2003
  • To enhance the performance of a serum-free medium (SFM) for human thrombopoietin (hTPO) production in suspension cultures of recombinant Chinese hamster ovary (rCHO) cells, several low-cost hydrolysates such as yeast hydrolysate (YH), soy hydrolysate, wheat gluten hydrolysate and rice hydrolysate were tested as medium additives. Among various hydrolysates tested, the positive effect of YH on hTPO production was most significant. When 5 g/L YH was added to SFM, the maximum hTPO concentration in batch culture was 40.41 ${\mu}g/mL$, which is 11.5 times higher than that in SFM without YH supplementation. This enhanced hTPO production in YH-supplemented SFM was obtained by the combined effect of enhanced $q_{hTPO}$ and increased culture longevity. In addition YH supplementation did not affect in vivo biological activity of hTPO. Taken together, the results obtained demonstrate the potential of YH as a medium additive for hTPO production in serum-free suspension cultures of rCHO cells.

  • PDF

Chinese Hamster Ovary세포에서 Benzo(a)pyrene과 3-Methylcholanthrene에 의한 DNA 단사절단과 복제억제에 미치는 Saponin의 영향

  • 이정섭;이형호;박기현;박상대
    • 한국환경성돌연변이발암원학회지
    • /
    • 제10권2호
    • /
    • pp.127-134
    • /
    • 1990
  • 본 연구는 Chinese hamster ovary (CHO-KI) 배양세포에서 S-15분획에 의해 활성화된 bezo(a) pyrene (BP)과 3-methylcholanthrene(MC)에 의해 유발된 DNA단사절단과 DNA복제억제 및 그 회복과정에 미치는 한국산 인삼추출물 saponin의 영향을 조사하여 다음과 같은 결과를 얻었다. S-15분획으로 활성화된 10-5 M의 BP 또는 MC를 1.0-10ng/ml의 saponin과 함께 처리할 경우 BP와 MC단독처리군에 비해 DNA 단사절단률이 감소하였다. DNA 합성률은 활성화된 10-5M의 BP와 10-6 M의 MC에 의해 각각 50% 및 75% 억제되었으나, 0.1-10ng/ml의 saponin을 동시에 처리할 경우 DNA 합성억제률이 약 10% 이상 둔화되었다.

  • PDF

Chinese Hamster Ovary세포에 있어 methyl methanesulfonate에 의한 DNA 복제억제와 이의 회복경로 (Replication Inhibition and Its Recovery/Process in Chinese Hamster Ovary Cells Treated with Methyl Methanesulfonate)

  • 이천복;이형호;박상대;이치건
    • 한국환경성돌연변이발암원학회지
    • /
    • 제9권1호
    • /
    • pp.33-46
    • /
    • 1989
  • 본 연구는 알킬화제를 처리한 CHO-K1 세포에서 DNA 복제억제와 그 회복과정의 분자론적 기작을 규명할 목적으로 방사선 이중 표지에 의한 DNA 합성율의 측정, 알칼리 자당 농도구배 초원심분리법에 의한 DNA 분자량과 후복제 회복율을 측정하여 다음과 같은 결과를 얻었다. (1) 1mM methyl methanesulfonate (MMS)와 1nM N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) 이하의 낮은 농도의 처리군에서는 DNA 합성율이 급격히 감소하였으나, 2 mM MMS, 2mM MNNG이상의 농도에서는 그 감소양상이 둔화되었다, (2) DNA 합성율은 알킬화제의 처리 직후 감소하였다가 시간경과에 따라 회복되어 처리후 4시간 째에는 대조군 수준 또는 그 이상으로 회복되었다.

  • PDF

SYNERGISTIC EFFECT OF HUMAN CYTOCHROME B5 COEXPRESSION ON THE METABOLIC ACTIVITY OF CYP1A2 IN CHINESE HAMSTER OVARY CELLS

  • Kang, Jin-Sun;Kang, Hyuck-Joon;Dong, Mi-Sook;Park, Chang-Hwan
    • 한국독성학회:학술대회논문집
    • /
    • 한국독성학회 2001년도 International Symposium on Dietary and Medicinal Antimutgens and Anticarcinogens
    • /
    • pp.188-188
    • /
    • 2001
  • Human cytochrome B5 (CYB5) was coexpressed with cytochrome P450 1A2 (CYP1A2), NADPH-CYP450 reductase (CYPR) and Ν-acetyltransferase 2 (NAT2) in Chinese hamster ovary (CHO) cells. The expression of four proteins was determined by Western blot analyses. The introduction of cDNAs to CHO cells were transduced via retroviral vectors. The cytotoxicity assay of 2-aminoanthracene (2-AA) and aflatoxin B$_1$were approximately 4-fold more sensitive than CYB5 free cells.(omitted)

  • PDF

SIMULTANEOUS EXPRESSION OF HUMAN CYTOCHROME P450 3A5 AND NADPH-CYTOCHROME P450 REDUCTASE IN CHINESE HAMSTER OVARY CELL USING INTERNAL RIBOSOME ENTRY SITE

  • Kang, Hyuck-Joon;Kang, Jin-Sun;Dong, Mi-Sook;Park, Chang-Hwan
    • 한국독성학회:학술대회논문집
    • /
    • 한국독성학회 2001년도 International Symposium on Dietary and Medicinal Antimutgens and Anticarcinogens
    • /
    • pp.189-189
    • /
    • 2001
  • For a continuous expression of human cytochrome p450 3A5 (CYP3A5) and NADPH-cytochrome P450 reductase (CYPR) proteins, bicistronic construct (CYP3A5BC-LNCX2) was made using internal ribosome entry site (IRES). As for mammalian cell expression, we used pLNCX2 retroviral vector; and using calcium phosphate, plasmid transfer was achieved in 293GPG cell and transduced in Chinese hamster ovary (CHO) cell.(omitted)

  • PDF

Chinese hamster ovary세포에서 발현된 pres2 및 S부위 함유 HBsAg의 항체유발능 (Antibody productivity of HBsAg containing both preS2 and S regions expressed in Chinese hamster ovary cells)

  • 정성균;박정민;이상봉;박동우;김동연;김기호;김홍진
    • 약학회지
    • /
    • 제45권6호
    • /
    • pp.708-714
    • /
    • 2001
  • Many studies have provided evidences that hepatitis B surface antigen (HBsAg) including preS region could be an ideal candidate for a new hepatitis B virus (HBV) vaccine with higher efficacy. We established CHO cell lines, IY-CHO-2 and IY-CHO-11 expressing high levels of HBsAg containing preS2 and S protein by stable transfection method. These cell lines expressed the correct size (about 1 kb in length) of HBsAg mRNA as expected. The purified protein from the culture supernatants of the clones showed the same sizes as those expressed in native hepatitis B virus (24 kDa, 27 kDa, 34 kDa and 36 kDa). Antibody productivity of CHO-derived HBsAg protein at lower dose challenge was higher than the protein containing S region alone expressed in yeast system. These results indicate that CHO-derived HBsAg protein containing preS2 and S region can be effectively used for a better immune response as a HBV vaccine.

  • PDF