Polydopamine and dopamine interfere with tetrazolium-based cytotoxicity assays and produce exaggerated cytocompatibility inferences

2021 ◽  
Author(s):  
Anand Kumar Awasthi ◽  
Sakshi Gupta ◽  
Kavthekar Rupesh Namdev ◽  
Aditi Banerjee ◽  
Aasheesh Srivastava
Keyword(s):  
Cell Viability ◽  
Trypan Blue ◽  
Reliable Estimate ◽  
Trypan Blue Exclusion ◽  
Cytotoxicity Assays ◽  
Trypan Blue Exclusion Assay

Polydopamine (PDA) and dopamine (DA) can spontaneously reduce MTT reagent to formazan, resulting in incorrect cell-viability inferences. The non-redox Trypan Blue exclusion assay provides a more reliable estimate of cell viability with PDA and DA.

scholarly journals Inhibitory Effects of Indirubin Derivatives on the Growth of HL-60 Leukemia Cells

2010 ◽  
Vol 5 (1) ◽  
pp. 1934578X1000500
Author(s):  
Nguyen Manh Cuong ◽  
Bui Huu Tai ◽  
Dang Hoang Hoan ◽  
Tran Thu Huong ◽  
Young Ho Kim ◽  
...  
Keyword(s):  
Cell Viability ◽  
Cancer Cells ◽  
Trypan Blue ◽  
Promyelocytic Leukemia ◽  
Leukemia Cells ◽  
Cytotoxic Activities ◽  
Inhibitory Effects ◽  
Trypan Blue Exclusion ◽  
Trypan Blue Exclusion Method ◽  
Exclusion Method

Six indirubin derivatives have been synthesized and their inhibitory effects on the growth of HL-60 human promyelocytic leukemia cells investigated. Cell viability was determined using the trypan blue exclusion method. Indirubin-3′-oxime (I-1) inhibited the growth of HL-60 cells with a GI50 value of 36.6 μM, whereas I-0, I-2, I-3, I-4 and I-6 showed only weak cytotoxic activities against HL-60 cancer cells with GI50 values in the range of 97.3 to over 100 μM. These results indicate that indirubin derivatives might be useful candidate agents for exploring potential antileukemic drugs.

scholarly journals Trypan Blue Exclusion Assay Verifies in Vitro Cytotoxicity of New Cis-Platinum (II) Complex in Human Cells

2019 ◽  
Vol 16 (3) ◽  
pp. 0555
Author(s):  
Hussein Et al.
Keyword(s):  
Cell Viability ◽  
Trypan Blue ◽  
Membrane Integrity ◽  
Platinum Complex ◽  
In Vitro Cytotoxicity ◽  
Toxic Effects ◽  
Polymorphonuclear Cells ◽  
Trypan Blue Exclusion ◽  
High Concentrations

          Various assays are used to determine the toxic effects of drugs at cellular levels in vitro.  One of these methods is the dye exclusion assay, which measures membrane integrity in the presence of Trypan blue. Trypan blue the dye which was used in this study to investigate cytotoxic effect of a new Cis –dichloroplatinum (II) complex [(Qu)2PtCl2] on the viability of polymorphonuclear cells (PMNs). Three concentrations of platinum complex were prepared (70, 35and 17.5 µg/ ml) and the results revealed that the percentage of cell viability decreased as the platinum complex concentration increased in comparison with control. The platinum complex exhibited low cytotoxic effects towards healthy cells at the concentrations of 17.5 µg/ ml and 35 µg/ ml, in which the percentage of cell viability was (77.01 ± 6.3) and (72.3± 0.50)respectively, with no significant differences as compared with the control(90.66 ±0.577). The viability was significantly decreased (67.59 ± 3.16) when the cells were treated with the concentration of 70 µg/ ml in comparison with control. These results indicated that the percentage of living cells decreased when treated with high concentrations of [(Qu)2PtCl2], which causes cells death, while low concentrations of the compound show low toxicity. This data indicates that this compound, at these concentrations may be suitable for use as a cancer treatment because it has low toxic effects on the healthy cells.        

scholarly journals Trypan blue exclusion assay by flow cytometry

2014 ◽  
Vol 47 (4) ◽  
pp. 307-315 ◽  
Author(s):  
B.A. Avelar-Freitas ◽  
V.G. Almeida ◽  
M.C.X. Pinto ◽  
F.A.G. Mourão ◽  
A.R. Massensini ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Trypan Blue ◽  
Trypan Blue Exclusion ◽  
Trypan Blue Exclusion Assay

Development of preovulatory follicles and oocytes during the oestrous cycle of mature and aged rats

1982 ◽  
Vol 100 (3) ◽  
pp. 434-443 ◽  
Author(s):  
J. J. Peluso ◽  
R. Hutz ◽  
C. England-Charlesworth
Keyword(s):  
Cell Viability ◽  
Granulosa Cells ◽  
Trypan Blue ◽  
Antral Follicle ◽  
Oestrous Cycle ◽  
Lower Percentage ◽  
Aged Rats ◽  
Trypan Blue Exclusion ◽  
Preovulatory Follicles ◽  
Lower Fertility

Abstract. The percentage of viable granulosa cells and the viability of the oocyte of each antral follicle on each day of the oestrous cycle of mature and aged rats was determined by trypan blue exclusion and fluorescein diacetate assays, respectively. In mature rats, the number of mid-sized follicles (350–550 μm in diameter) increased from oestrus through di-oestrus. The percentage of viable granulosa cells increased in only mid-sized follicles which contained a viable oocyte. On pro-oestrus, two types of preovulatory follicles (> 550 μm in diameter) were observed in the mature pro-oestrous rat: those with a high granulosa cell viability (> 60%) and a viable oocyte and those with a lower percentage of viable granulosa cells (< 50%) and a non-viable oocyte. This suggests that in mature rats only viable oocytes would be ovulated since presumably, only those preovulatory follicles with a high percentage of viable granulosa cells are able to ovulate. In aged rats, preovulatory-sized follicles were present on each day of the cycle. The percentage of viable granulosa cells within those preovulatory-sized follicles with nonviable oocytes increased on met-oestrus, decreased on di-oestrus and increased by pro-oestrus. This pattern resulted in the development of 3 to 4 preovulatory follicles per ovary with all preovulatory-sized follicles possessing a high percentage of viable granulosa cells regardless of the functional state of the oocyte. It is proposed that in aged rats, all the preovulatory follicles would ovulate but many of the ovulated oocytes would be defective. This mechanism would account for the lower fertility of these older animals.

Clarification of the Phenotypic Characteristics and Anti-Tumor Activity of Hedyotis diffusa

2011 ◽  
Vol 39 (01) ◽  
pp. 201-213 ◽  
Author(s):  
Hong-Zin Lee ◽  
Da-Tian Bau ◽  
Chao-Lin Kuo ◽  
Ru-Yin Tsai ◽  
Yu-Chang Chen ◽  
...  
Keyword(s):  
Trypan Blue ◽  
High Performance ◽  
Significant Inhibition ◽  
Lung Cancer Cell ◽  
Phenotypic Characteristics ◽  
Trypan Blue Exclusion ◽  
Trypan Blue Exclusion Assay ◽  
Hedyotis Diffusa ◽  
Anti Tumor Activity ◽  
Hplc Data

Hedyotis diffusa Willd. (Rubiaceae) is an important folk herb used to prevent and cure hepatitis and liver cancer in Taiwan. For differentiation of H. diffusa from counterfeits, macroscopic and microscopic characters of H. diffusa, H. corymbosa and H. tenelliflora were examined in this study. According to Trypan blue exclusion assay and Western blot analysis, H. diffusa had a significant inhibition of cell growth and induction of cell apoptosis in COLO 205 (colon cancer), Hep 3B (hepatocellular carcinoma) and H460 (lung cancer) cell lines. This study also used high-performance liquid chromatography (HPLC) to determine the quality control of H. diffusa. The HPLC data showed that ursolic and oleanolic acid are the components of the H. diffusa, consisting of approximately 4.66–4.80% and 1.86–1.96%, respectively. Our study also demonstrated that ursolic acid has significant anti-tumor activity in COLO 205, Hep 3B and H460 cancer cells.

scholarly journals Induction of Necrosis and DNA Fragmentation during Hypothermic Preservation of Hepatocytes in UW, HTK, and Celsior Solutions

2003 ◽  
Vol 12 (1) ◽  
pp. 59-68 ◽  
Author(s):  
Salomon L. Abrahamse ◽  
Pieter Van Runnard Heimel ◽  
Robin J. Hartman ◽  
Rob A. F. M. Chamuleau ◽  
Thomas M. Van Gulik
Keyword(s):  
Cell Death ◽  
Cell Viability ◽  
Dna Fragmentation ◽  
Trypan Blue ◽  
Caspase 3 ◽  
Trypan Blue Exclusion ◽  
Hypothermic Preservation ◽  
Mtt Reduction ◽  
Ldh Release ◽  
Porcine Hepatocytes

Donor cells can be preserved in University of Wisconsin (UW), histidine-tryptophan-ketoglutarate (HTK), or Celsior solution. However, differences in efficacy and mode of action in preventing hypothermia-induced cell injury have not been unequivocally clarified. Therefore, we investigated and compared necrotic and apoptotic cell death of freshly isolated primary porcine hepatocytes after hypothermic preservation in UW, HTK, and Celsior solutions and subsequent normothermic culturing. Hepatocytes were isolated from porcine livers, divided in fractions, and hypothermically (4°C) stored in phosphate-buffered saline (PBS), UW, HTK, or Celsior solution. Cell necrosis and apoptosis were assessed after 24- and 48-h hypothermic storage and after 24-h normothermic culturing following the hypothermic preservation periods. Necrosis was assessed by trypan blue exclusion, lactate dehydrogenase (LDH) release, and mitochondrial 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) reduction. Apoptosis was assessed by the induction of histone-associated DNA fragments and cellular caspase-3 activity. Trypan blue exclusion, LDH release, and MTT reduction of hypothermically preserved hepatocytes showed a decrease in cell viability of more than 50% during the first 24 h of hypothermic preservation. Cell viability was further decreased after 48-h preservation. DNA fragmentation was slightly enhanced in hepatocytes after preservation in all solutions, but caspase-3 activity was not significantly increased in these cells. Normothermic culturing of hypothermically preserved cells further decreased cell viability as assessed by LDH release and MTT reduction. Normothermic culturing of hypothermically preserved hepatocytes induced DNA fragmentation, but caspase-3 activity was not enhanced in these cells. Trypan blue exclusion, LDH leakage, and MTT reduction demonstrated the highest cell viability after storage in Celsior, and DNA fragmentation was the lowest in cells that had been stored in PBS and UW solutions. None of the preservation solutions tested in this study was capable of adequately preventing cell death of isolated porcine hepatocytes after 24-h hypothermic preservation and subsequent 24-h normothermic culturing. Culturing of isolated and hypothermically preserved hepatocytes induces DNA fragmentation, but does not lead to caspase-3 activation. With respect to necrosis and DNA fragmentation of hypothermically preserved cells, UW and Celsior were superior to PBS and HTK solutions in this model of isolated porcine hepatocyte preservation.

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