• 제목/요약/키워드: Cell Viability

검색결과 3,686건 처리시간 0.031초

벼 원형질체의 분리, 배양 및 Electroporation에 관한 연구 (Isolation, Culture and Electroporation of Rice Protoplasts)

  • 황성진
    • Journal of Plant Biology
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    • 제34권1호
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    • pp.19-23
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    • 1991
  • 벼의 종자로부터 embryogenic callus를 유도하고 이로부터 개체로의 재분화를 유도하였으며, embroyogenic cell suspension culture를 통하여, 이로부터 원형질체를 분리, 배양하여 embryogenic callus를 형성하였다. 또한, 분리된 원형질체를 electroporation하였을 때 생존율(viability)에 미치는 여러 요인들을 조사하였다. 원형질체의 생존율은 voltage와 capactiance가 증가할수록 감소를 보였으며, HBM buffer에서 $4^{\circ}C$로 electroporation 하였을 때 생존율에 보다 효과적이었다.

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Induction of Apoptotic Cell Death by Red Pericarp Rice (Jakwangchalbyeo) Extracts

  • Chi, Hee-Youn;Lee, Chang-Ho;Kim, Kwang-Ho;Kim, Sun-Lim;Chung, Ill-Min
    • Food Science and Biotechnology
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    • 제15권4호
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    • pp.534-542
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    • 2006
  • The effects of ethanol fractions of three different rice grain extracts, Jakwangchalbyeo, Hwasunchalbyeo, and Ilpumbyeo, on apoptotic cell death in the rat hepatoma H4IIE cell line were investigated using the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] cell viability assay. One hundred mg/mL Jakwangchalbyeo extract significantly reduced cell viability to 69.5, 57.2, and 46.1% within 24, 48, and 72 hr, respectively. Fluorescence-activated cell sorting (FACS) analyses were also performed to characterize the cell death pattern caused by treatment with the rice grain extracts. Apoptotic cell death was clearly observed with time after treatment with the Jakwangchalbyeo extract. In Western blotting analysis, degradation of the 116 kDa poly-ADP-ribose polymerase (PARP) molecule was observed with concomitant formation of an 89 kDa product 24, 48, and 72 hr after treating cells with the Jakwangchalbyeo extract. This indicates that an apoptotic process caused cell death in these cells. In conclusion, red-pericarp Jakwangchalbyeo extract induced apoptotic cell death in H4IIE cells to a larger extent than the other rice extracts.

The Effect of NMDA/glycine Receptor Antagonist, 7-Chlorokynurenic Acid on Cultured Astrocytes Damaged by Ischemia-like Condition

  • 정인주
    • 대한의생명과학회지
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    • 제15권4호
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    • pp.355-362
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    • 2009
  • I evaluated the protective effect of N-methyl-D-aspartate (NMDA)/glycine receptor antagonist, 7-chlorokinurenic acid (CKA) on cultured mouse astrocytes damaged by ischemia-like condition (ILC). The protective effect of CKA was assessed by cell viability, lactate dehydrogenase (LDH) activity, superoxide dismutase (SOD)-like activity and lipid peroxidation. To examine the effect of CKA on the cell apoptosis, the expression and the activity of caspase 3 were assessed by Western blotting. CKA increased the cell viability decreased by ILC. CKA also decreased the LDH activity and antioxidative effects such as SOD-like activity and inhibitory activity of lipid peroxidation. In addition, CKA suppressed the expression of caspase 3 associated with apoptosis, and increased the cell viability by the decrease of caspase 3 activity as like the caspase 3 inhibitor, Av-DVED-MED. From these results, these results suggest that ILS induces cell cytotoxicity in cultured astrocytes and CKA, NMDA/glycine receptor antagonist, is effective on the prevention of the cytotoxicity due to ILS by the antioxidative effect and the inhibition of apoptosis.

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형질전환된 Nicotiana tabacum 배양에 있어서 glutathione과 ascorbic acid가 세포생장과 생존율에 미치는 영향

  • 김용훈;이상윤;김동일
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2003년도 생물공학의 동향(XII)
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    • pp.293-297
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    • 2003
  • 본 연구에서는 형질전환된 N. tabacum 배양에 있어서 glutathione과 ascorbic acid가 세포 생장과 생존율에 미치는 영향을 비교하여 실험하였다. Glutathione과 ascorbic acid의 첨가는배양초기 세포 생존율의 감소를 완화시켰으며, 그로인한 재조합 단백질의 생산성 증대를 확인할 수 있었다. 또한 glutathione과 ascorbic acid를 첨가하여 배양한 세포는 cold stress로 인한 세포 생존율의 감소와 DNA 절편화를 억제 시키는데 효과가 있었다. 변형시킨 FDA법에 의한 세포 생존율은 배양 6일째 최대를 보였으며, 고 삼투압 배지와 저온의 환경은 세포 생존율을 저하시켰다. 반면 cold stress 전에 glutathione과 ascorbic acid를 첨가하여 배양한 세포의 생존율은 cold stress 후, 대조구 세포보다 높게 유지되었으며, DNA 절편화 현상도 cold stress 후, 대조구 세포보다 적게 일어남을 확인하였다. 따라서 glutathione과 ascorbic acid는 cold stress로 의한 세포 생존율 저하를 완화시키며, apoptosis 억제 효과가 있다고 판단된다.

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Protective Effect of Crataegi Fructus Extract on the Neurotoxicity Induced by Reactive Oxygen Species in Cultured C6 Glioma Cell

  • ;유선미
    • 대한의생명과학회지
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    • 제14권1호
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    • pp.27-32
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    • 2008
  • To clerify the antioxidant effect of Crataegi Fructus (CF) extract on reactive oxygen species (ROS), The C6 glioma cells were treated with various concentrations of hydrogen peroxide ($H_2O_2$). The $H_2O_2$-induced neurotoxicity was measured by XTT assay for the cell viability. For the protective effect of CF extract on the cytotoxicity induced by $H_2O_2$, cell viability, lactate dehydroganase (LDH) activity, and the inhibitive activity of lipid peroxidation of CF extract were performed. In this study, $H_2O_2$ decreased cell viability dose- and time-dependent manners and increased LDH activity compared with the control. In the protective effect on $H_2O_2$, CF extract increased cell viability and decreased LDH activity on $H_2O_2$-induced cytotoxicity, lipid peroxidation by FTC assay. From these results, It is suggested that $H_2O_2$ was highly toxic on cultured C6 glioma cells, and also, CF extract showed the protective effect on $H_2O_2$-mediated cytotoxicity.

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대두 Protoplast의 세포벽 합성과 세포분열에 대한 Cytokinin의 영향 (Effects of Cytokinin on Cell Wall Regeneration and Cell Division of Soybean Protoplasts)

  • 류기중;김형옥;박창규;김창오
    • Applied Biological Chemistry
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    • 제30권4호
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    • pp.300-304
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    • 1987
  • 대두(Glycine max var. Acme)의 자엽조직에서 유래된 callus의 부유배양세포로부터 protoplast를 분리하고, 이들 protoplast의 viability, 세포벽재생 및 세포분열에 대한 benzyladenine(BA)의 영향과 protoplast에 의한 BA의 흡수특성을 조사하였다. Protoplast의 viability는 BA처리에 의하여 증가되었고, 세포벽재생과 세포분열 그리고 callue의 생장도 BA처리에 의하여 촉진되었다. Protoplast에 의하여 흡수된 BA의 양은 BA 처리후 약 20시간에 최대에 이르렀고, 이중에서 약 2/3가 6시간 이내에 흡수되었다.

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허혈이 유도된 대뇌신경세포에 대한 항산화제 및 Ampa/kainate 수용체 길항제의 영향 (Effect of Antioxidant and Ampa/kainate Receptor Antagonist on Cerebral Neurons Damaged by Ischemia)

  • 오연균
    • 동의생리병리학회지
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    • 제19권4호
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    • pp.1022-1026
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    • 2005
  • To clarify the toxic effect on cultured neonatal mouse cerebral neurons damaged by ischemia, we examined the cytotoxicity induced by ischemia and the protective effect of antioxidant and AMPA/kainate receptor antagonist against ischemia-induced cytotoxicity on cultured cerebral neurons. For this study, mice were administrated with 20ug/kg cyclothiazide or 50U/kg vitamin E via intraperitoneal injection for 2 hours before ischemic induction. After cell culture for 7 days, cell viability, amount of neurofilament and protein kinase C activity were examined. Ischemia decreased significantly cell viability, amount of neurofilament and the increase of protein kinase C activity in these cultures. In the protective effect, vitamin I showed remarkably the increase of cell viability and amount of neurofilament, and the decrease of protein kinase C activity but, cyclothiazide did not showed any protective effect on ischemia-induced cytotoxicity. From these results, it is suggested that vitamin I is effective in blocking the neurotoxicity induced by ischemia, but cyclothiazide as a AMPA/kainate receptor antagonist is not.

생간건비탕(生肝健脾湯)이 HepG2 cell의 증식, 세포사멸 및 활성조절 신호전달계에 미치는 영향 (The Effects of Saengkankunbi-tang on Proliferation, Apoptosis and Cell Signaling Pathways of HepG2 Cells)

  • 김재용;김영철;이장훈;우홍정
    • 대한한방내과학회지
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    • 제27권1호
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    • pp.149-165
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    • 2006
  • Objectives: This study was done to evaluate the effects of Saengkankunbi-tang on cell-viability, proliferation, cell-cycle, apoptosis and DNA replication on HepG2 cell and to find out by which molecular-biological mechanism by which Saengkankunbi-tang operates. Methods : The MTT assay, cell counting assay, [3H]-thymidine incorporation assay, flow cytometric analysis, tryphan blue exclusion assay, western blot analysis, quantative RT-PCR were taken. Results : Saengkankunbi-tang had no effect on proliferation, cell-cycle and DNA replications of HepG2 cells, while it improved cell viability and reduced apoptosis, and it activated Akt and NFKB. But, it did not produce an effect on cell viability and apoptosis when P13K/Akt pathway was blocked by LY294002 nor when $NF{\kapa}B$ activation was blocked by DN-$I{\kapa}B$. Conclusion : These results suggests that Saengkankunbi-tang improves cell viability and reduces apoptosis of HepG2 cells, by activating $NF{\kapa}B$ through PI3K/Akt pathway.

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Mori Fructus Induces Cell Death through ROS-dependent Mitochondrial Apoptotic Pathway in Human Glioma Cells

  • Jang, Sang-Won;Jeong, Ji-Cheon
    • 동의생리병리학회지
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    • 제22권5호
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    • pp.1322-1329
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    • 2008
  • Mulberry has been reported to contain wide range of polyphenols and have chemopreventive activity. However, little has been known regarding the effect of mulberry fruits on cell viability in human glioma cells. The present study was undertaken to examine the effect of mulberry fruit (Mar; Fructus) on cell viability and to determine its underlying mechanism in human glioma cells. Cell viability and cell death were estimated by MTT assay and trypanblue exclusion assay, respectively. Reactive oxygen species (ROS) generation was measured using the fluorescence probe DCFH-DA. The mitochondrial transmembrane potential was measured with $DiOC_6$(3). Bax expression and cytochrome c release were measured by Western blot analysis. Caspase activity was estimated using colorimetric kit. Mori Fructus resulted in apoptotic cell death in a dose- and time-dependent manner. Mori Fructus increased ROS generation and the Mori Fructus-induced cell death was also prevented by antioxidants, suggesting that ROS generation plays a critical role in Mari Fructus-induced cell death. Western blot analysis showed that Mori Fructus treatment caused an increase in Bax expression, which was inhibited by the antioxidant N-acetylcysteine (NAC). Mori Fructus induced depolarization of mitochondrial membrane potential and its effect was inhibited by the antioxidants NAC and catalase. Mori Fructus induced cytochrome c release, which was inhibited by NAC. Caspase activity was stimulated by Mori Fructus and caspase inhibitors prevented the Mori Fructus-induced cell death. These findings suggest that Mori Fructus results in human glioma cell death through ROS-dependent mitochondrial pathway in human glioma cells.

Lactobacillus acidophilus Contributes to a Healthy Environment for Vaginal Epithelial Cells

  • Pi, Woo-Jin;Ryu, Jae-Sook;Roh, Jae-Sook
    • Parasites, Hosts and Diseases
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    • 제49권3호
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    • pp.295-298
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    • 2011
  • Lactobacillus species in the female genital tract are thought to act as a barrier to infection. Several studies have demonstrated that lactobacilli can adhere to vaginal epithelial cells. However, little is known about how the adherence of lactobacilli to vaginal epithelial cells affects the acidity, cell viability, or proliferation of the lactobacilli themselves or those of vaginal epithelial cells. Lactobacillus acidophilus was co-cultured with immortalized human vaginal epithelial cells (MS74 cell line), and the growth of L. acidophilus and the acidity of the culture medium were measured. MS74 cell density and viability were also assessed by counting cell numbers and observing the cell attachment state. L. acidophilus showed exponential growth for the first 6 hr until 9 hr, and the pH was maintained close to 4.0-5.0 at 24 hr after culture, consistent with previous studies. The growth curve of L. acidophilus or the pH values were relatively unaffected by co-culture with MS74 cells, confirming that L. acidophilus maintains a low pH in the presence of MS74 cells. This co-culture model could therefore potentially be used to mimic vaginal conditions for future in vitro studies. On the other hand, MS74 cells co-cultured with L. acidophilus more firmly attached to the culture plate, and a higher number of cells were present compared to cells cultured in the absence of L. acidophilus. These results indicate that L. acidophilus increases MS74 cell proliferation and viability, suggesting that lactobacilli may contribute to the healthy environment for vaginal epithelial cells.