Cell Culture for Testing Anticancer Compounds

Ruben, Regina L.

doi:10.1016/b978-0-12-007906-3.50012-7

Cited by 10 publications

(9 citation statements)

References 145 publications

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“…Advantages: This method is simple, inexpensive, and a good indicator of membrane integrity [17], and dead cells are colored blue within seconds of exposure to the dye [18].…”

Section: Trypan Blue Dye Exclusion Assaymentioning

confidence: 99%

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In Vitro Cytotoxicity and Cell Viability Assays: Principles, Advantages, and Disadvantages

Aslantürk¹

2018

Genotoxicity - A Predictable Risk to Our Actual World

275

241

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Cytotoxicity is one of the most important indicators for biological evaluation in vitro studies. In vitro, chemicals such as drugs and pesticides have different cytotoxicity mechanisms such as destruction of cell membranes, prevention of protein synthesis, irreversible binding to receptors etc. In order to determine the cell death caused by these damages, there is a need for cheap, reliable and reproducible short-term cytotoxicity and cell viability assays. Cytotoxicity and cell viability assays are based on various cell functions. A broad spectrum of cytotoxicity assays is currently used in the fields of toxicology and pharmacology. There are different classifications for these assays: (i) dye exclusion assays; (ii) colorimetric assays; (iii) fluorometric assays; and (iv) luminometric assays. Choosing the appropriate method among these assays is important for obtaining accurate and reliable results. When selecting the cytotoxicity and cell viability assays to be used in the study, different parameters have to be considered such as the availability in the laboratory where the study is to be performed, test compounds, detection mechanism, specificity, and sensitivity. In this chapter, information will be given about in vitro cytotoxicity and viability assays, these assays will be classified and their advantages and disadvantages will be emphasized. The aim of this chapter is to guide the researcher interested in this subject to select the appropriate assay for their study.

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“…Advantages: This method is simple, inexpensive, and a good indicator of membrane integrity [17], and dead cells are colored blue within seconds of exposure to the dye [18].…”

Section: Trypan Blue Dye Exclusion Assaymentioning

confidence: 99%

“…Therefore, counting errors (~10%) could be occurred. Counting errors have been attributed to poor dispersion of cells, cell loss during cell dispersion, inaccurate dilution of cells, improper filling of the chamber and presence of air bubbles in the chamber [17].…”

Section: Trypan Blue Dye Exclusion Assaymentioning

confidence: 99%

In Vitro Cytotoxicity and Cell Viability Assays: Principles, Advantages, and Disadvantages

Aslantürk¹

2018

Genotoxicity - A Predictable Risk to Our Actual World

275

241

View full text Add to dashboard Cite

show abstract

“…metric tetrazolium reduction assay, designed to detect both cytostatic and cytocidal drug effects [20,21]. Optimal cell seeding concentrations were predetermined to maximize dye reduction over a 96 hr incubation period [21].…”

Section: Tetrazolium Reduction Assay (Mtt Assay)mentioning

confidence: 99%

“…Optimal cell seeding concentrations were predetermined to maximize dye reduction over a 96 hr incubation period [21]. Concentrations ranged from 1.25 x 103 cells/well for B16F10 cells to 2 x 105 cells/well for HPBMC and spleen cells.…”

Section: Tetrazolium Reduction Assay (Mtt Assay)mentioning

confidence: 99%

“…A clonogenic assay on a solid, hydrophilic substrate [21] was used to distinguish between cytostatic and cytocidal effects of ptilocaulin in MCF-7 and B16F10 cell lines in which the compound was a potent growth inhibitor (IC50 _ 0.1 ffg/ml in the MTT assay). Cell growth inhibition in the presence of compound followed by resumption of growth in the absence of compound was considered a cytostatic effect.…”

Section: Clonogenic Assaymentioning

confidence: 99%

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Cytotoxicity of synthetic racemic ptilocaulin: A novel cyclic guanidine

Ruben¹,

Snider²,

Hobbs³

et al. 1989

Invest New Drugs

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(+)-Ptilocaulin, a novel cyclic guanidine extracted from the Caribbean sponge Ptilocaulis aff. P. Spiculifer, is reported to have broad spectrum antimicrobial activity in vitro as well as in vitro activity against L1210 murine leukemia. To more fully evaluate this compound as an anticancer agent, the in vitro cell growth inhibitory potencies of synthetic racemic ptilocaulin and ten clinical anticancer drugs were determined and compared in 16 different normal and transformed human and murine cell populations. Potency, expressed as the 50% inhibitory concentration (IC50), was determined by a tetrazolium reduction (MTT) assay. Ptilocaulin showed a fairly broad spectrum of in vitro activity against colon and mammary adenocarcinomas, melanomas, leukemias, transformed fibroblasts and normal lymphoid cells (IC50s 0.05- greater than 10 micrograms/ml). This activity was comparable to that of many of the clinical drugs, including vinca alkyloids, antibiotics, alkylators and antimetabolites. Cell viability was affected only after a 72 hr exposure to the compound. In a clonogenic assay, cytocidal effects were observed after 24-72 hr exposures to 10 x IC50 concentrations of ptilocaulin, as evidenced by failure of cells to resume growth after removal of the compound. Cytostatic effects were observed at less than or equal to IC50 concentrations, as evidenced by resumption of growth to near-control levels after removal of the compound. Ptilocaulin was toxic at 50 and 25 mg/kg in an in vivo L1210 tumor model and was ineffective at lower concentrations (T/Cs 100-112%). In vivo studies in a more sensitive tumor system are recommended but are limited by the lack of availability of sufficient quantities of the compound.

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Cytotoxic Activity of Herbal Medicines as Assessed in Vitro: A Review

Gavanji

Bakhtari

Famurewa

et al. 2023

Chemistry & Biodiversity

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Since time immemorial, human beings have sought natural medications for treatment of various diseases. Weighty evidence demonstrates the use of chemical methodologies for sensitive evaluation of cytotoxic potentials of herbal agents. However, due to the ubiquitous use of cytotoxicity methods, there is a need for providing updated guidance for the design and development of in vitro assessment. The aim of this review is to provide practical guidance on common cell-based assays for suitable assessment of cytotoxicity potential of herbal medicines and discussing their advantages and disadvantages Relevant articles in authentic databases, including PubMed, Web of Science, Science Direct, Scopus, Google Scholar and SID, from 1950 to 2022 were collected according to selection criteria of in vitro cytotoxicity assays and protocols. In addition, the link between cytotoxicity assay selection and different factors such as the drug solvent, concentration and exposure duration were discussed.

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Cell Culture for Testing Anticancer Compounds

Cited by 10 publications

References 145 publications

In Vitro Cytotoxicity and Cell Viability Assays: Principles, Advantages, and Disadvantages

In Vitro Cytotoxicity and Cell Viability Assays: Principles, Advantages, and Disadvantages

Cytotoxicity of synthetic racemic ptilocaulin: A novel cyclic guanidine

Cytotoxic Activity of Herbal Medicines as Assessed in Vitro: A Review

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