• Title/Summary/Keyword: cell line

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Taurine Transporter Activity in the Human Colon Carcinoma cell Line(HT-29) is Decreased during Cell Differentiation (인체 소장상피세포주(HT-29)의 분화단계에 따른 타우린수송체 활성의 변화)

  • 박태선
    • Journal of Nutrition and Health
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    • v.33 no.6
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    • pp.660-667
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    • 2000
  • Previous studies on the effect of age on intestinal taurine transport in animals have invariably shown a decline in the activity of the transport system with increasing age. In the present study changes in taurine transporter activity were observed during cell differentiation in the human colon carcinoma cell line HT-29 This cell line exhibits various enterocytic characteristics when differentiated and therefore has frequently been used to study the characteristcs and regulation of nutrient and drug absorption in the small intestine at the cellular level. Pre-treatment of the cells with $\beta$-alanine(10mM) reduced the taurine transport activity to 33% of the value for the control cells(p<0.05) which implies that taurine and $\beta$-alanine share a common $\beta$-amino acid transport system for their celluar uptake in the HI-29 was continued until 21 days post seeding. Kinetic studies of the taurine transporter were conducted in the HT-29 cell line with varying taurine concentration(5-60$\mu$M) in the uptake medium Both Vmax and the Michaelis-Menten constant(Km) of taurine transporter were decreased as differentiation of the HT-29 cell line was progressed ; Vmax of the taurine transporter in cells incubated for 4, 14 and 21 days post seeding was 2.79$\pm$3.4m 16.89$\pm$1.74, and 0.85$\pm$0.08 and 0.32$\pm$0.01nmol.mg protein-1 .30min-1 respectively(p<0.001) and Km was 42.3$\pm$3.4, 16.89$\pm$1.74, and 11.2$\pm$3.0$\mu$M respectively (p<0.01) These results indicate that the activity of sodium dependent active taurine transport system in the HT-29 cell line is decreased as confluent cells are differentiated. This phenomenon in cell culture system corresponds well with the earlier observation of lower intestinal taurine transport activity in suckling rats compared to that in adult animals although direct relationship of cell differentiation with in vivo aging process needs further verification.

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Studies on toxicity of ochratoxin A to chromosomes of turmor cell-line (종양세포(腫瘍細胞)의 염색체(染色體)에 대한 오크라톡신 A의 독성(毒性)에 관한 연구(硏究))

  • Yoon, Wha-jung;Roh, Min-hee;Kim, Kang-ryun
    • Korean Journal of Veterinary Research
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    • v.29 no.2
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    • pp.51-57
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    • 1989
  • This study was performed to investigate the toxicity of ochratoxin A (OA) to the chromosomes of $K_{562}$ tumor cell-line in vitro. The results of this experiment were as follows: 1) Chromosomes of $K_{562}$tumor cell-line resulted in pseudotriploidy on the control group. Chromosomes of $K_{562}$ tumor cell-line treated with OA resulted in heteroploidy compared with the control group. The mean number of chromosomes in the karyotype of the control group (60) were 7 in the A group, 5 in the B group, 20 in the C+X group, 7 in the D group, 9 in the E group, 6 in the F group, and 6 in the G+Y group respectively. The number of chromosomes were increased as follows: Treating with $0.7{\mu}M$ OA, the number of chromosomes were increased one in E and F group, two in G+Y group compared with control group. In treated with $1.5{\mu}M$ OA, the increasing number of chromosome was one in E and F group. In treated with $3{\mu}M$ OA, E and F group was increased one and G+Y group were increased two chromosomes compared with control group. But in treated with $6{\mu}M$ OA, the number of chromosome in G+Y group was decreased one. 2) $K_{562}$ tumor cell line treated with OA showed Philadelphia-Chromosome in the long arm of the G group karyotype chromosome. The rate of chromosome aberration in $K_{562}$ tumor cell-line treated with OA was 77% in $0.7{\mu}M$ OA group, 71% in $1.5{\mu}M$ OA group, 82% in $3{\mu}M$ OA group and 94% in $6{\mu}M$ OA group respectively. The rate of chromosome aberration of $K_{562}$ tumor cell-line treated with OA was high in the high dose level of OA, and chromosome aberration of $K_{562}$ tumor cell-line treated with OA showed deletion, minute, dicentric-chromosome and translocation in the long arm of the C-group karyotype. As a result of this study, the toxicity of OA showed deletion, minute, dicentric-chromosome and translocation in the long arm of the C-group karyotype, and then, the toxicity of OA resulted in the damage to RNA and protein synthesis in $K_{562}$ tumor cell-line, and the C-group karyotype of $K_{562}$ tumor cell-line was target of the toxicity of OA.

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Reduction of Ammonia Accumulation and Improvement of Cell Viability by Expression of Urea Cycle Enzymes in Chinese Hamster Ovary Cells

  • Chung, Myung-Il;Lim, Mi-Hee;Lee, Yun-Jeong;Kim, Ik-Hwan;Kim, Ick-Young;Kim, Jung-Hoe;Chang, Kern-Hee;Kim, Hong-Jin
    • Journal of Microbiology and Biotechnology
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    • v.13 no.2
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    • pp.217-224
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    • 2003
  • Previously, we developed a CHO cell line (CHO-OTC1-Al9) that expresses the first two enzymes in the urea cycle and exhibits a higher ammonia-removing ability and faster growth rate than a vector-controlled CHO cell line (CHO-neo-5). The current study was undertaken to develop a cell line with an ammonia-removing ability higher than the cell line developed previously. To accomplish this, CHO cell lines expressing the first three, first four, or all five enzymes of the urea cycle were constructed using a stable transfection method. Finally, the CHO-AS-16, CHO-AL-19, and CHO-Arg-11 cell lines expressing the first three, first four, and all five enzymes of the urea cycle, respectively, were selected and found to exhibit higher ammonia-removing ability than the CHO-OTC1-Al9 cell line. Among the three selected cell lines, CHO-AL-19 showed the highest ammonia-removing ability and highest cell viability at a higher cell density, with 40% and 15% lower ammonia concentration in the, culture media than that of CHO-neo-5 and CHO-OTC1-A19 cell lines, respectively. CHO-AL-19 also showed 44% and 10% higher cell viability than the CHO-neo-5 and CHO-OTC-Al9 cell lines, at a higher cell density, respectively. The ammonia concentrations in the culture media were expressed as the ammonia concentration/cell, and the CHO-AL-19 cells revealed 45-60% and 20% lower ammonia concentration/cell than the CHO-neo-5 and CHO-OTC1-Al9 cells, respectively.

Comparative Characterization of Growth and Recombinant Protein Production among Three Insect Cell Lines with Four Kinds of Serum Free media

  • Kwon, Mi-Sun;Takashi Dojima;Park, Enoch Y.
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.8 no.2
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    • pp.142-146
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    • 2003
  • Three insect cell lines, Sf9, Sf21 and Tn5Bl-4, and four different kinds of serum free media (SFM), Sf 900 II, EX-CELL 420, EX-CELL 405 and Express Five, were used to compare the nutrient consumption, byproduct formation, production of recombinant protein and protease activity in suspension cultures. The Sf 900 II SFM was a ppropriate for the cell growth and protein production of the Sf9 and Sf21 cell lines. When the Tn5Bl-4 cell line was grown in the Express Five SFM, the specific growth rate was 1.6 fold higher than those of either the Sf9 or Sf21 cell lines. The glucose and glutamine consumption rates per cells, were 4 and 2.3 times higher than those of the Sf9 cell line, respectively. The overall yield coefficients of the lactate and ammoniumion were 2.8 and 1.5 times higher compared to those of the Sf9 cell line. respectively. The maximum specific ${\beta}$-galactosidase production rate was 4.5 fold that of the Sf9 cell line, a 3 times higher protease activity per cell.

Effects of Carbamoyl Phosphate Synthetase I against Cell Growth and Production of Recombinant Erythropoietin in Urea Cycle Enzyme Expressing CHO Cell Line (Carbamoyl Phosphate Synthetase I이 요소회로 유전자를 발현하는 CHO 세포 주의 세포 성장과 재조합 Erythropoietin의 생산에 미치는 영향)

  • Cho, Su-Mi;Kim, Na-Young;Kim, Hyoung-Jin;Kim, Hong-Jin
    • YAKHAK HOEJI
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    • v.51 no.3
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    • pp.214-218
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    • 2007
  • In the previous reports, we developed the CO5 by introducing genes for the first and second urea cycle enzymes, carbamoyl phosphate synthetase I (CPS I) and ornithine transcarbamoylase (OTC) into the IBE cell lines producing erythropoietin (EPO). The CO5 have been found out to have 15-20% higher cell growth rate and produce 2-times more EPO than the parental cell line, IBE. To investigate the role of CPS I in CO5 cell line for the cell growth and amount of EPO, we knock-downed CPS I gene expression via siRNA treatment. Expression level of EPO in cell lysate of CO5 was 3-5 fold higher than that of IBE. After siRNA treatment, the cell growth of CO5 was decreased 8-21% and the EPO productivity in the cell Iysate was significantly decreased. However, these changes of the cell growth and EPO productivity were not observed in IBE. These results indicate that CPS I gene expression is important for the increased cell growth and EPO productivity of CO5 cell line.

Microcarrier Culture of an Anchorage-dependent Cell Using Cytodex-3 (Cytodex-3를 이용한 부착성 동물세포의 미립담체 배양)

  • 김정회;최준호;웨이슈후
    • Microbiology and Biotechnology Letters
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    • v.17 no.3
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    • pp.231-235
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    • 1989
  • Possibility of using microcarriers for the growth of a transformed human embryonic kidney cell line 293 was investigated. The cell grew well in a static culture such as T-flasks with medium of DME/F12 (3:1) mixture supplemented with 5% FBS, but it was most difficult to make the cells grow on microcarriers mainly due to the low attachment efficiency and poor spreading at initial stage of the culture. Consequently, 30-50% of the cells were lost upon inoculation into microcarrier suspension and significant fraction of the mirrocarrier became bald. The medium supplemented with the concentrated conditioned medium by hepatoma cell line HpG2 supported the active growth of the cells on microcarrier and the cells showed a very healthy and well spreading morphology. It was probable that some spreading and attachment factors of HpG2 conditioned medium were effective for 293 cells.

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The Effects of KakamBoyangHwanoh-Tang(KBHT) and PalMihapChongMung-Tang(PMCMT) on Protecting Microglia and Inhibiting Acetylcholinesterase and Oxidants (가미보양환오탕(加味補陽還五湯)과 팔미합총명탕(八味合聰明湯)의 microglia 보호, 항산화 및 acetylcholinesterase 억제 효과)

  • Kim, Hyun-Joo;Lee, Sang-Ryong
    • Journal of Oriental Neuropsychiatry
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    • v.19 no.2
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    • pp.65-75
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    • 2008
  • Objective : This experiment was designed to investigate the effect of the KBHT and PMCMT extract on protecting microglia and inhibiting acetylcholinesterase and oxidants. Method : The effects of the KBHT and PMCMT extract on cell death of BV2 microglial cell line treated by ${\gamma}$ ; expression of NO, ROS in BV2 microglial cell line treated by lipopolysaccharide(LPS) ; AChE activity in PC-12 cell treated by NGF were investigated, respectively. Result : The KBHT and PMCMT extract significantly increased cell viability in BV2 microglial cell line treated with ${\gamma}$. The KBHT and PMCMT extract suppressed the NO and ROS production in BV2 microglial cell line treated by LPS. The KBHT and PMCMT extract groups also showed inhibition of AChE activity in PC-12 cell line. Conclusion : According to the above result, it is suggested that the KBHT and PMCMT extract might be usefully applied for prevention and treatment of Alzheimer' s disease.

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Effects of OnDam-TanghapChongMyoung-Tang and DoDam-TanghapChongMyoung-Tang on Protecting Microglia and Inhibiting Acetylcholinesterase and Oxidants (온담탕합총명탕(溫膽湯合聰明湯)과 도담탕합총명탕(導痰湯合聰明湯)의 microglia 보호, 항산화 및 acetylcholinesterase 억제효과)

  • Cheong, Myong-Hee;Jung, In-Chul
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.5
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    • pp.1276-1282
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    • 2008
  • This experiment was designed to investigate the effect of the ODTCMT and DDTCMT extract on protecting microglia and inhibiting acetylcholinesterase and oxidants. The effects of the ODTCMT and DDTCMT extract on cell death of BV2 microglial cell line treated by $IFN-{\gamma}$ ; expression of NO, ROS in BV2 microglial cell line treated by lipopolysaccharide (LPS) ; AChE activity in PC-12 cell treated by NGF were investigated, respectively. The ODTCMT and DDTCMT extract significantly increased cell viability in BV2 microglial cell line treated with $IFN-\nu$. The ODTCMT and DDTCMT extract suppressed the NO and RDS production in BV2 microglial cell line treated by LPS. The ODTCMT and DDTCMT extract groups also showed inhibition of AChE activity in PC-12 cell line. According to the above result, it is suggested that the ODTCMT and DDTCMT extract might be usefully applied for prevention and treatment of Alzheimer's disease. OnDam-TanghapChongMyoung-Tang (ODTCMT), DoDam-TanghapChongMyoung-Tang (DDTCMT), Microglia, acetylcholinesterase, ROS

Effects of ChenwhangBosim-Dan and SungsimJihwang-Tang on Protecting Microglia and Inhibiting Acetylcholinesterase and Oxidants (천왕보심단(天王補心丹)과 성심지황탕(醒心地黃湯)의 microglia 보호, 항산화 및 acetylcholinesterase 억제 효과)

  • Jung, In-Chul
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.1
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    • pp.120-125
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    • 2008
  • This experiment was designed to investigate the effect of the CBD and SJT extract on protecting microglia and inhibiting acetylcholinesterase and oxidants. The effects of the CBD and SJT extract on cell death of BV2 microglial cell line treated by $IFN-{\gamma}$ ; expression of NO, ROS in BV2 microglial cell line treated by lipopolysaccharide(LPS) ; AChE activity in PC-12 cell treated by NGF were investigated, respectively. The CBD and SJT extract significantly increased cell viability in BV2 microglial cell line treated with $IFN-{\gamma}$. The CBD and SJT extract suppressed the NO and ROS production in BV2 microglial cell line treated by LPS. The CBD and SJT extract groups also showed inhibition of AChE activity in PC-12 cell line. According to the above result, it is suggested that the CBD and SJT extract might be usefully applied for prevention and treatment of Alzheimer's disease.

The effects of Honey Bee Venom for Aqua-acupuncture on Expression of Genes Related with Inflammation and Pain (봉독(蜂毒) 약침액(藥鍼液)이 염증(炎症) 및 통증(痛症) 관련(關聯) 유전자(遺傳子) 발현(發現)에 미치는 영향(影響))

  • Jeong, Hye-Yoon;Koh, Hyung-Kyun
    • Journal of Acupuncture Research
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    • v.19 no.3
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    • pp.41-50
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    • 2002
  • Objective : To study anti-inflammatory, analgesic effect and molecular biological mechanism of honey bee venom for aqua-acupuncture, human mast cell line(HMC-1) and human glioma cell line(HS683) were treated with bee venom. Methods : Cell viability of bee venom was tested by 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) asssay. To explore whether anti-inflammatory, analgesic effects of bee venom are associated with the control of gene expression, quantitative RT-PCR analysis of inflammation and pain related genes was performed. Results : The MTT assay demonstrated that cell viability was not decreased by treatment with 10-9 ug/ml bee venom in comparison with 10-2, 10-3, 10-4, 10-5, 10-6, 10-7, 10-8, 10-9, 10-10 and 10-11 ug/ml. sPLA2 and COX-l were down-regulated by treatment with 10-9 ug/ml bee venom in HS683 Cell line in comparison with control. COX-2 was up-regulated by treatment with 10-9 ug/ml bee venom in HS683 Cell line and HSP-2 was up-regulated by treatment with 10-9 ug/ml bee venom in HMC-1 Cell line in comparison with control. sPLA2, COX-1 and COX-2 showed no significant regulation in HMC-1 Cell line and cPLA2 also showed no significant regulation in both HMC-l and HS683 Cell line between control and bee venom treated group.

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