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Screening of compounds toxicity against human Monocytic cell line-THP-1 by flow cytometry


The worldwide rapid increase in bacterial resistance to numerous antibiotics requires on-going development of new drugs to enter the market. As the development of new antibiotics is lengthy and costly, early monitoring of compound’s toxicity is essential in the development of novel agents. Our interest is in a rapid, simple, high throughput screening method to assess cytotoxicity induced by potential agents. Some intracellular pathogens, such as Mycobacterium tuberculosis primary site of infection is human alveolar macrophages. Thus, evaluation of candidate drugs for macrophage toxicity is crucial. Protocols for high throughput drug toxicity screening of macrophages using flow cytometry are lacking in the literature. For this application we modified a preexisting technique, propidium iodide (PI) exclusion staining and utilized it for rapid toxicity tests. Samples were prepared in 96 well plates and analyzed by flow cytometry, which allowed for rapid, inexpensive and precise assessment of compound’s toxicity associated with cell death.



Fluorescence Activated Cell Sorting


propidium iodide

LD50 :

lethal dose in which 50% of the cells die


  1. Tufts Center for the Study of Drug Development. Impact report 4(5) Sept 2002.

  2. Tufts Center for the Study of Drug Development. Impact report 5(3) May 2003.

  3. Alliance pharmaceutical corp: phases of product development. Drug development and approval process; p. 1–3.

  4. Dye C, Scheele S, Dolin P, Pathania V, Raviglione MC. Consensus statement. Global burden of tuberculosis: estimated incidence, prevalence, and mortality by Country. WHO Global Surveillance and Monitoring Project, JAMA 1999; 282(7):677–686.

    Article  PubMed  CAS  Google Scholar 

  5. WHO/IUATLD. Global project on anti-tuberculosis drug resistance in the world: Second report. World Health Organization/CDS/2000.278.

  6. Carryn S, Van Bambeke F, Mingeot-Leclercq MP, Tulkens PM. Comparative intracellular (THP-1 Macrophage) and extracellular activities of -lactams, Azithromycin, Gentamicin and Fluoroquinolones against Listeria monocytogenes at clinically relevant concentrations. Anti Agent Chemo 2002; 46(7):2095–2103.

    Article  CAS  Google Scholar 

  7. Takemura H, Hiroyuki Yamamato H, Kunishima H, Ikejima H, Takashi H et al. Evaluation of a human monocytic cell line THP-1 model for assay of the intracellular activities of antimicrobial agents against Legionella pneumophila. J Antimicrob Chemother 2000; 46(4):589–594.

    Article  PubMed  CAS  Google Scholar 

  8. Darzynkiewicz Z, Bruno S, Del Bino G, Gorczyca W, Hotz MA, Lassota P, Traganos F. Features of Apoptotic Cells Measured by Flow Cytometry. Cytometry 1992; 13: 795- 808.

    Article  PubMed  CAS  Google Scholar 

  9. Pick N, Good RL, Arad D, Av-Gay Y. EXEG 1706: A novel antimicrobial agent against Methicillin-resistant Staphylococcus aureus. Abs- F-2147 (pp. 269) ICAAC, Chicago 2003.

    Google Scholar 

  10. Vermes I, Haanen C, Steffens-Nakken H, Reutelingsperger C. A novel assay for apoptosis. Flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labeled Annexin V. J Immunol Methods 1995;184:39–51.

    Article  PubMed  CAS  Google Scholar 

  11. Schmid I, Krall WJ, Uittenbogaart CH, Braun J, Giorgi JV. Dead cell discrimination with 7-amino-actinomycin D in combination with dual color immunofluorescence in single laser flow cytometry. Cytometry 1992; 13(2):204–208.

    Article  PubMed  CAS  Google Scholar 

  12. Darzynkiewicz, Z, Juan G, Li X, Gorezvca W, Murakami T, Traganos F. Cytometry in cell necrobiology: analysis of apoptosis and accidental cell death (necrosis). Cytometry 1997; 27(1):1–20.

    Article  PubMed  CAS  Google Scholar 

  13. Sherwood SW, Schimke RT. Cell cycle analysis of apoptosis using flow cytometry. Methods Cell Biol 1995; 46:77–97.

    Article  PubMed  CAS  Google Scholar 

  14. Telford WG, King LE, Fraker PJ. Comparative evaluation of several DNA binding dyes in the detection of apoptosisassociation chromatin degradation by flow cytometry. Cytometry 1992; 13(2):137–143.

    Article  PubMed  CAS  Google Scholar 

  15. Bertho AL, Santiago MA, Coutinho SG. Flow cytometry in the study of cell death. Mem Inst Oswaldo Cruz 2000; 95(3):429–433.

    PubMed  CAS  Google Scholar 

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Correspondence to Yossef Av-Gay.

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Published: October 1, 2004.

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Pick, N., Cameron, S., Arad, D. et al. Screening of compounds toxicity against human Monocytic cell line-THP-1 by flow cytometry. Biol. Proced. Online 6, 220–225 (2004).

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Indexing terms

  • Flow Cytometry
  • Propidium
  • Lethal Dose 50