Skip to main content
SpringerLink
Log in

Differential Staining Cytotoxicity Assay: A Review

  • Protocol
  • First Online:
Book cover Cancer Cell Culture

Part of the book series: Methods in Molecular Biology ((MIMB,volume 731))

Abstract

Differential Staining Cytotoxicity (DiSC) assay is the prototype for a closely related family of assays based on the concept of total cell kill, or, in other words, cell death occurring in the entire population of tumor cells. It is probably the most versatile of the cell-death end points, in that it (1) can be applied to both solid and hematologic neoplasms, (2) can be applied to specimens in which it is not possible to obtain a pure population of highly enriched tumor cells, and (3) can be applied to a wide variety of drugs, ranging from traditional cytotoxic agents to biological response modifiers with activity mediated through tumor-infiltrating effector cells, to “targeted” kinase inhibitors, and to antivascular agents, such as bevacizumab and pazopanib. The basic principles of the assay are to culture three-dimensional fresh tumor cell clusters in anchorage-independent conditions. At the conclusion of the culture period, Fast Green dye is added to the microwells, the contents of which are then sedimented onto permanent Cytospin centrifuge slides and then counterstained with hematoxylin–eosin or Wright–Giemsa. “Living” cells stain with the cytologic stain in question and can be identified as either normal or neoplastic, based on standard morphologic criteria. “Dead” cells stain blue-green. Nonviable endothelial cells appear as strikingly hyperchromatic, blue-black, and often refractile objects, which may be readily distinguished from other types of dead cells. This assay has been biologically and clinically validated in a number of ways, as described in this chapter.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 139.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 179.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 249.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Weisenthal, L.M., Shoemaker, R.H., Marsden, J.A., Dill, P.L., Baker, J.A. and Moran, E.M. (1984) In vitro chemosensitivity assay based on the concept of total tumor cell kill. Recent Results Cancer Res. 94, 161–73.

    Article  PubMed  CAS  Google Scholar 

  2. Weisenthal, L.M. and Lippman, M.E. (1985) Clonogenic and nonclonogenic in vitro chemosensitivity assays. Cancer Treat. Rep. 69, 615–32.

    PubMed  CAS  Google Scholar 

  3. Weisenthal, L.M. (1993) Cell culture assays for hematologic neoplasms based on the concept of total tumor cell kill. In: Drug Resistance in Leukemia and Lymphoma, G.J.L. Kaspers, Pieters, R., Twentyman, P.R., Weisenthal, L.M., and Veerman, A.J.P., Editors., Har-wood Academic Publishers: Langhorne, PA. p. 415–432.

    Google Scholar 

  4. Weisenthal, L.M. (1994) Clinical correlations for cell culture assays based on the concept of total tumor cell kill. Contrib. Gynecol. Obstet. 19, 82–90.

    PubMed  CAS  Google Scholar 

  5. Bosanquet, A.G., Nygren, P., and Weisenthal, L.M. (2008) Individualized tumor response testing in leukemia and lymphoma. In: Innovative Leukemia and Lymphoma Therapy, G.J.L. Kaspers, Coiffier, B., Heinrich,M.C., and Estey,E.H., Editors, Informa Healthcare: New York. p. 23–43.

    Google Scholar 

  6. Weisenthal, L.M. and Nygren, P. (2002) Current Status of Cell Culture Drug Resistance Testing (CCDRT). http://weisenthal.org/oncol_t.htm.

  7. Hickman, J.A. (1992) Apoptosis induced by anticancer drugs. Cancer Metastasis Rev. 11, 121–39.

    Article  PubMed  CAS  Google Scholar 

  8. Zunino, F., Perego, P., Pilotti, S., Pratesi, G., Supino, R. and Arcamone, F. (1997) Role of apoptotic response in cellular resistance to cytotoxic agents. Pharmacol. Ther. 76, 177–85.

    Article  PubMed  CAS  Google Scholar 

  9. Jaffrezou, J.P., Bettaïeb, A., Levade, T. and Laurent, G. (1998) Antitumor agent-induced apoptosis in myeloid leukemia cells: a controlled suicide. Leuk. Lymphoma. 29, 453–63.

    Article  PubMed  CAS  Google Scholar 

  10. Castejon, R., Yebra, M., Citores, M.J., Villarreal, M., García-Marco, J.A. and Vargas, J.A. (2009) Drug induction apoptosis assay as predictive value of chemotherapy response in patients with B-cell chronic lymphocytic leukemia. Leuk. Lymphoma 50, 593–603.

    Article  PubMed  CAS  Google Scholar 

  11. Weisenthal, L.M., Marsden, J.A., Dill, P.L. and Macaluso, C.K. (1983) A novel dye exclusion method for testing in vitro chemosensitivity of human tumors. Cancer Res. 43, 749–57.

    PubMed  CAS  Google Scholar 

  12. Weisenthal, L.M., Dill, P.L., Finklestein, J.Z., Duarte, T.E., Baker, J.A. and Moran, E.M. (1986) Laboratory detection of primary and acquired drug resistance in human lymphatic neoplasms. Cancer Treat. Rep. 70, 1283–95.

    PubMed  CAS  Google Scholar 

  13. Carmichael, J., DeGraff, W.G., Gazdar, A.F., Minna, J.D. and Mitchell, J.B. (1987) Evaluation of a tetrazolium-based semiautomated colorimetric assay: assessment of chemosensitivity testing. Cancer Res. 47, 936–42.

    PubMed  CAS  Google Scholar 

  14. Kangas, L., Gronroos, M. and Nieminen A.L. (1984) Bioluminescence of cellular ATP: a new method for evaluating cytotoxic agents in vitro. Med. Biol. 62, 338–43.

    PubMed  CAS  Google Scholar 

  15. Garewal, H.S., Ahmann, F.R., Schifman, R.B. and Celniker, A. (1986) ATP assay: ability to distinguish cytostatic from cytocidal anticancer drug effects. J. Natl. Cancer Inst. 77, 1039–45.

    PubMed  CAS  Google Scholar 

  16. Sevin, B.U., Peng, Z.L., Perras, J.P., Ganjei, P., Penalver, M. and Averette, H.E. (1988) Application of an ATP-bioluminescence assay in human tumor chemosensitivity testing. Gynecol. Oncol. 31, 191–204.

    Article  PubMed  CAS  Google Scholar 

  17. Rotman, B., Teplitz, C., Dickinson, K. and Cozzolino, J.P. (1988) Individual human tumors in short-term micro-organ cultures: chemosensitivity testing by fluorescent cytoprinting. In Vitro Cell. Dev. Biol. 24, 1137–46.

    Article  PubMed  CAS  Google Scholar 

  18. Larsson, R., Nygren, P., Ekberg, M. and Slater, L. (1990) Chemotherapeutic drug sensitivity testing of human leukemia cells in vitro using a semiautomated fluorometric assay. Leukemia 4, 567–71.

    PubMed  CAS  Google Scholar 

  19. Nygren, P., Kristensen, J., Jonsson, B., Sundström, C., Lönnerholm, G., Kreuger, A. and Larsson, R. (1992) Feasibility of the fluorometric microculture cytotoxicity assay (FMCA) for cytotoxic drug sensitivity testing of tumor cells from patients with acute lymphoblastic leukemia. Leukemia 6, 1121–8.

    PubMed  CAS  Google Scholar 

  20. Twentyman, P.R., Fox, N.E. and Rees, J.K. (1989) Chemosensitivity testing of fresh leukaemia cells using the MTT colorimetric assay. Br. J. Haematol. 71,19–24.

    Article  PubMed  CAS  Google Scholar 

  21. Pieters, R., Huismans, D.R., Leyva, A. and Veerman, A.J. (1989) Comparison of the rapid automated MTT-assay with a dye exclusion assay for chemosensitivity testing in childhood leukaemia. Br. J. Cancer 59, 217–20.

    Article  PubMed  CAS  Google Scholar 

  22. Pieters, R., Huismans, D.R., Leyva, A. and Veerman, A.J. (1989) Sensitivity to purine analogues in childhood leukemia assessed by the automated MTT-assay. Adv. Exp. Med. Biol. 253A, 447–54.

    PubMed  CAS  Google Scholar 

  23. Kirkpatrick, D.L., Duke, M. and Goh, T.S. (1990) Chemosensitivity testing of fresh human leukemia cells using both a dye exclusion assay and a tetrazolium dye (MTT) assay. Leuk. Res. 14, 459–66.

    Article  PubMed  CAS  Google Scholar 

  24. Tsai, C.M., Ihde, D.C., Kadoyama, C., Venzon, D. and Gazdar, A.F. (1990) Correlation of in vitro drug sensitivity testing of long-term small cell lung cancer cell lines with response and survival. Eur. J. Cancer 26, 1148–52.

    Article  PubMed  CAS  Google Scholar 

  25. Dmitrovsky, E., Seifter, E.J., Gazdar, A.F., Tsai, C.M., Edison, M., Brantley, P., Veach, S.R., Batist, G., Ihde, D.C. and Mulshine, J.L. (1990) A phase II trial of carboplatin (CBDCA) in small-cell and non-small-cell lung cancer with correlation to in vitro analysis of cytotoxicity. Am. J. Clin. Oncol. 13, 285–9.

    Article  PubMed  CAS  Google Scholar 

  26. Hanson, J.A., Bentley, D.P., Bean, E.A., Nute, S.R. and Moore, J.L. (1991) In vitro chemosensitivity testing in chronic lymphocytic leukaemia patients. Leuk. Res. 15, 565–9.

    Article  PubMed  CAS  Google Scholar 

  27. Rhedin, A.S., Tidefelt, U., Jönsson, K., Lundin, A. and Paul, C. (1993) Comparison of a bioluminescence assay with differential staining cytotoxicity for cytostatic drug testing in vitro in human leukemic cells. Leuk. Res. 17, 271–6.

    Article  PubMed  CAS  Google Scholar 

  28. Nygren, P., Hagberg, H., Glimelius, B., Sundström, C., Kristensen, J., Christiansen, I. and Larsson, R. (1994) In vitro drug sensitivity testing of tumor cells from patients with non-Hodgkin’s lymphoma using the fluorometric microculture cytotoxicity assay. Ann. Oncol. 5 Suppl 1, 127–31.

    Google Scholar 

  29. Weisenthal, L.M. and Marsden, J.A. (1981) A novel dye exclusion assay for predicting response to cancer chemotherapy. Proc. Am. Assoc. Cancer Res. 22, 155.

    Google Scholar 

  30. Weisenthal, L., Marsden, J.A., Malefatto, J. and Dill, P.L. (1981) Predicting response to cancer chemotherapy with novel dye exclusion assay. In: XIIth International Congress of Chemotherapy. Florence, Italy.

    Google Scholar 

  31. Carmichael, J., DeGraff, W.G., Gazdar, A.F., Minna, J.D. and Mitchell, J.B. (1987) Evaluation of a tetrazolium-based semiautomated colorimetric assay: assessment of radiosensitivity. Cancer Res. 47, 943–6.

    PubMed  CAS  Google Scholar 

  32. Pappenheimer, A.M. (1917) Experimental Studies Upon Lymphocytes: I. The Reactions of Lymphocytes under Various Experimental Conditions. J. Exp. Med. 25, 633–650.

    Google Scholar 

  33. Richter, M.N. and Macdowell, E.C. (1933) Studies on Mouse Leukemia: Vii. The Relation of Cell Death to the Potency of Inoculated Cell Suspensions. J. Exp. Med. 57, 1–20.

    Google Scholar 

  34. Schrek, R. (1936) A method for counting the viable cells in normal and in malignant cell suspensions. Am. J. Cancer 28, 389–392.

    Google Scholar 

  35. Schrek, R. and Ott, J.N.Jr. (1952) Study of the death of irradiated and non-irradiated cells by time-lapse cinemicrography. AMA Arch. Pathol. 53, 363–78.

    PubMed  CAS  Google Scholar 

  36. Vycital, R.O., Schrek, R. and Clarke, T.H. (1953) Unstained cell counts as a method of evaluating cancerocidal agents. J. Lab. Clin. Med. 42, 326–34.

    PubMed  CAS  Google Scholar 

  37. Schrek, R., Leithold, S.L. and Friedman, I.A. (1957) In vitro sensitivity of human leukemic cells to x-rays. Proc. Soc. Exp. Biol. Med. 94, 250–3.

    PubMed  CAS  Google Scholar 

  38. Schrek, R., Friedman, I.A. and Leithold, L. (1958) Variability of the in vitro sensitivity of human leukemic lymphocytes to x-rays and chemotherapeutic agents. J. Natl. Cancer. Inst. 20, 1037–50.

    PubMed  CAS  Google Scholar 

  39. Schrek, R., Leithold, S.L., Friedman, I.A. and Best, W.R. (1962) Clinical evaluation of an in vitro test for radiosensitivity of leukemic lymphocytes. Blood 20, 432–42.

    PubMed  CAS  Google Scholar 

  40. Schrek, R. (1964) Prednisolone Sensitivity and Cytology of Viable Lymphocytes as Tests for Chronic Lymphocytic Leukemia. J. Natl. Cancer Inst. 33, 837–47.

    PubMed  CAS  Google Scholar 

  41. Schrek, R. (1965) In vitro methods for measuring viability and vitality of lymphocytes exposed to 45 degree, 47 degree, and 50 degree C. Cryobiology 2, 122–8.

    Article  PubMed  CAS  Google Scholar 

  42. Dolowy, W.C., Elrod, L.M., Ammeraal, R.N. and Schrek, R. (1967) Toxicity of l-asparaginase to resistant and susceptible lymphoma cells in vitro. Proc. Soc. Exp. Biol. Med. 125, 598–601.

    PubMed  CAS  Google Scholar 

  43. Schrek, R. and Dolowy, W.C. (1971) In vitro test for sensitivity of leukemic cells to l-asparaginase. Cancer Res. 31, 523–6.

    PubMed  CAS  Google Scholar 

  44. Schrek, R. (1975) Sensitivity of leukaemic lymphocytes to microtubular reagents. Br. J. Exp. Pathol. 56, 280–5.

    PubMed  CAS  Google Scholar 

  45. Schrek, R. and Stefani, S.S. (1976) Cytarabine: cytocidal effect on normal and leukemic lymphocytes. Synergism with x-rays and comparison with mechlorethamine. Exp. Mol. Pathol. 24, 84–90.

    Google Scholar 

  46. Knospe, W.H., Gregory, S.A., Trobaugh, F.E. Jr., Stedronsky, J.A. and Schrek, R. (1977) Chronic lymphocytic leukemia: correlation of clinical course and therapeutic response with in vitro testing and morphology of lymphocytes. Am. J. Hematol. 2, 73–101.

    Article  PubMed  CAS  Google Scholar 

  47. Schrek, R. (1979) Utility and efficiency of viable cell counts. Cancer Res. 39, 4288.

    PubMed  CAS  Google Scholar 

  48. Schrek, R. and Stefani, S.S. (1980) Effects of hyperthermia on radiosensitivity of normal and leukaemic lymphocytes. Br. J. Exp. Pathol. 61, 256–60.

    PubMed  CAS  Google Scholar 

  49. Schrek, R. and Stefani, S.S. (1981) Toxicity of microtubular drugs to leukemic lymphocytes. Exp. Mol. Pathol. 34, 369–78.

    Article  PubMed  CAS  Google Scholar 

  50. Schrek, R., (1988) Chronic lymphocytic leukemic patients, resistant to chemotherapy. Med. Hypotheses 26, 227–8.

    Article  PubMed  CAS  Google Scholar 

  51. Schrek, R., Best, W.R. and Stefani, S. (1988) Relationship between in vitro and in vivo radiosensitivity of lymphocytes in chronic lymphocytic leukemia. Acta Haematol. 80, 129–33.

    Article  PubMed  CAS  Google Scholar 

  52. Schrek, R. (1988) Intractable chronic lymphocytic leukemia. J. Natl. Cancer. Inst. 80, 604.

    Google Scholar 

  53. Schrek, R. (1990) Essential in vitro test before treatment of patients with intractable chronic lymphocytic leukemia. Acta Haematol. 84, 104–5.

    Article  PubMed  CAS  Google Scholar 

  54. Schrek, R. (1990) Differences between responsive and intractable chronic lymphocytic leukemia. Med. Hypotheses 31, 81–2.

    Article  PubMed  CAS  Google Scholar 

  55. Schrek, R. (1991) Intractable chronic lymphocytic leukemia and interferon. Med. Hypotheses. 35, 182–3.

    Article  PubMed  CAS  Google Scholar 

  56. Durkin, W.J., Ghanta, V.K., Balch, C.M., Davis, D.W. and Hiramoto, R.N. (1979) A methodological approach to the prediction of anticancer drug effect in humans. Cancer Res. 39, 402–7.

    PubMed  CAS  Google Scholar 

  57. Salmon, S.E., Hamburger, A.W., Soehnlen, B., Durie, B.G., Alberts, D.S. and Moon, T.E. (1978) Quantitation of differential sensitivity of human-tumor stem cells to anticancer drugs. N. Engl. J. Med. 298, 1321–7.

    Article  PubMed  CAS  Google Scholar 

  58. Weisenthal, L.M., Dill, P.L., Kurnick, N.B. and Lippman, M.E. (1983) Comparison of dye exclusion assays with a clonogenic assay in the determination of drug-induced cytotoxicity. Cancer Res. 43, 258–64.

    PubMed  CAS  Google Scholar 

  59. Weisenthal, L.M., Lalude, A.O. and Miller J.B. (1983) In vitro chemosensitivity of human bladder cancer. Cancer 51, 1490–6.

    Article  PubMed  CAS  Google Scholar 

  60. Wilbur, D.W., Camacho, E.S., Hilliard, D.A., Dill, P.L. and Weisenthal, L.M. (1992) Chemotherapy of non-small cell lung carcinoma guided by an in vitro drug resistance assay measuring total tumour cell kill. Br. J. Cancer 65, 27–32.

    Article  PubMed  CAS  Google Scholar 

  61. Weisenthal, L.M., (1991) Predictive assays for drug and radiation resistance. In: Human Cancer in Primary Culture: A Handbook, J.M. Masters, Editor. Kluwer Academic Publishers: Dordrecht, The Netherlands.

    Google Scholar 

  62. Weisenthal, L.M., Dill, P. and Birkhofer, M. (1991) Accurate identification of disease-specific activity of antineoplastic agents with an in vitro fresh tumor assay measuring killing of largely non-dividing cells. Proc. Am. Assoc. Cancer Res. 32, 384.

    Google Scholar 

  63. Weisenthal, L.M. and Dill, P. (1992) In vitro effects of interleukin-2 (IL2) on fresh human tumor cell cultures measured by the DISC assay. Proc. Am. Assoc. Cancer Res. 33, A3313.

    Google Scholar 

  64. Kurohara, W., Colman, M., Nagourney, R.A., Weisenthal, L.M., Swingle, K. and Redpath, J.L. (1989) Radiation response of cells from human tumor biopsies as assessed by a dye exclusion technique: a possible predictive assay. Int. J. Radiat. Biol. 56, 767–70.

    Article  PubMed  CAS  Google Scholar 

  65. Weisenthal, L.M., Dill, P.L., and Swingle, K.F. (1989) Clinical radiation sensitivity profiles of human neoplasms are reproduced by a short-term in vitro (DiSC) assay measuring cytotoxicity in the total (largely nondividing) tumor cell population following ultra- high dose, single-fraction radiation. Proc. Am. Assoc. Cancer Res. 30, 401.

    Google Scholar 

  66. Weisenthal, C.L., Meade, R.C., Owenby, J. and Irwin, R.I. (1961) An investigation of kutapressin as a hemostatic agent in transurethral surgery of the prostate. J. Urol. 86, 346–9.

    PubMed  CAS  Google Scholar 

  67. Weisenthal, L.M., Su, Y.Z., Duarte, T.E. and Nagourney, R.A. (1988) Non-clonogenic, in vitro assays for predicting sensitivity to cancer chemotherapy. Prog. Clin. Biol. Res. 276, 75–92.

    PubMed  CAS  Google Scholar 

  68. Weisenthal, L.M., Nagourney, R.A., Kern, D.H., Boullier, B., Bosanquet, A.G., Dill, and M. P.L., J.C., and Moran, E.M. (1989) Approach to the clinical circumvention of drug resistance utilizing a non-clonogenic in vitro assay measuring the effects of drugs, radiation, and interleukin-II on largely non-dividing cells. In: Strategies in Cancer Medical Therapy: Biological Bases and Clinical Implications. Pavia, Italy: Edimes.

    Google Scholar 

  69. Weisenthal, L.M., Su, Y.Z., Duarte, T.E., Dill, P.L. and Nagourney, R.A. (1987) Perturbation of in vitro drug resistance in human lymphatic neoplasms by combinations of putative inhibitors of protein kinase C. Cancer Treat. Rep. 71, 1239–43.

    PubMed  CAS  Google Scholar 

  70. Moran, E., Nagourney, R.A., Ottenheimer, E.J., Mahutte, K., and Weisenthal, L.M. (1988) Reversal of acquired drug resistance with lidocaine and verapamil: a phase I study. Proc. Am. Assoc. Cancer Res. 29, 218.

    Google Scholar 

  71. Weisenthal, L.M. (1991) Effect of prior chemotherapy on biologic response modifier activity. J. Natl. Cancer Inst. 83, 790–3.

    Article  PubMed  CAS  Google Scholar 

  72. Weisenthal, L.M., Dill, P.L. and Pearson, F.C. (1991) Effect of prior cancer chemotherapy on human tumor-specific cytotoxicity in vitro in response to immunopotentiating biologic response modifiers. J. Natl. Cancer Inst. 83, 37–42.

    Article  PubMed  CAS  Google Scholar 

  73. Bosanquet, A.G., Bird, M.C., Price, W.J. and Gilby, E.D. (1983) An assessment of a short-term tumour chemosensitivity assay in chronic lymphocytic leukaemia. Br. J. Cancer 47, 781–9.

    Article  PubMed  CAS  Google Scholar 

  74. Bird, M.C., Bosanquet, A.G. and Gilby, E.D. (1985) In vitro determination of tumour chemosensitivity in haematological malignancies. Hematol. Oncol. 3, 1–10.

    Article  PubMed  CAS  Google Scholar 

  75. Bird, M.C., Bosanquet, A.G., Forskitt, S. and Gilby, E.D. (1986) Semi-micro adaptation of a 4–day differential staining cytotoxicity (DiSC) assay for determining the in-vitro chemosensitivity of haematological malignancies. Leuk. Res. 10, 445–9.

    Article  PubMed  CAS  Google Scholar 

  76. Bird, M.C., Godwin, V.A., Antrobus, J.H. and Bosanquet, A.G. (1987) Comparison of in vitro drug sensitivity by the differential staining cytotoxicity (DiSC) and colony-forming assays. Br. J. Cancer 55, 429–31.

    Article  PubMed  CAS  Google Scholar 

  77. Bird, M.C., Bosanquet, A.G., Forskitt, S. and Gilby, E.D. (1988) Long-term comparison of results of a drug sensitivity assay in vitro with patient response in lymphatic neoplasms. Cancer 61, 1104–9.

    Article  PubMed  CAS  Google Scholar 

  78. Bosanquet, A.G. (1991) Correlations between therapeutic response of leukaemias and in-vitro drug-sensitivity assay. Lancet 337, 711–4.

    Article  PubMed  CAS  Google Scholar 

  79. Bosanquet, A.G. (1993) In vitro drug sensitivity testing for the individual patient: an ideal adjunct to current methods of treatment choice. Clin. Oncol. (R. Coll. Radiol.) 5, 195–7.

    Article  CAS  Google Scholar 

  80. Bosanquet, A.G., McCann, S.R., Crotty, G.M., Mills, M.J. and Catovsky, D. (1995) Methylprednisolone in advanced chronic lymphocytic leukaemia: rationale for, and effectiveness of treatment suggested by DiSC assay. Acta Haematol. 93, 73–9.

    Article  PubMed  CAS  Google Scholar 

  81. Bosanquet, A.G. and Bell, P.B. (1996) Enhanced ex vivo drug sensitivity testing of chronic lymphocytic leukaemia using refined DiSC assay methodology. Leuk. Res. 20, 143–53.

    Article  PubMed  CAS  Google Scholar 

  82. Bosanquet, A.G., et al. 1997 Ex vivo cytotoxic drug evaluation by DiSC assay to expedite identification of clinical targets: results with 8-chloro-cAMP. Br J Cancer 76(4): p. 511–8.

    Article  PubMed  CAS  Google Scholar 

  83. Bosanquet, A.G., Copplestone, J.A., Johnson, S.A., Smith, A.G., Povey, S.J., Orchard, J.A. and Oscier, D.G. (1999) Response to cladribine in previously treated patients with chronic lymphocytic leukaemia identified by ex vivo assessment of drug sensitivity by DiSC assay. Br. J. Haematol. 106, 474–6.

    Article  PubMed  CAS  Google Scholar 

  84. Bosanquet, A.G., Johnson, S.A. and Richards, S.M. (1999) Prognosis for fludarabine therapy of chronic lymphocytic leukaemia based on ex vivo drug response by DiSC assay. Br. J. Haematol. 106, 71–7.

    Article  PubMed  CAS  Google Scholar 

  85. Mason, J.M., Drummond, M.F., Bosanquet, A.G. and Sheldon, T.A. (1999) The DiSC assay. A cost-effective guide to treatment for chronic lymphocytic leukemia? Int. J. Technol. Assess. Health Care 15, 173–84.

    Google Scholar 

  86. Bosanquet, A.G. and Bosanquet, M.I. (2000) Ex vivo assessment of drug response by differential staining cytotoxicity (DiSC) assay suggests a biological basis for equality of chemotherapy irrespective of age for patients with chronic lymphocytic leukaemia. Leukemia 14, 712–5.

    Article  PubMed  CAS  Google Scholar 

  87. Bosanquet, A.G., Burlton, A.R. and Bell, P.B. (2002) Parameters affecting the ex vivo cytotoxic drug sensitivity of human hematopoietic cells. J. Exp. Ther. Oncol. 2, 53–63.

    Article  PubMed  CAS  Google Scholar 

  88. Thornton, P.D., Matutes, E., Bosanquet, A.G., Lakhani, A.K., Grech, H., Ropner, J.E., Joshi, R., Mackie, P.H., Douglas, I.D., Bowcock, S.J. and Catovsky, D. (2003) High dose methylprednisolone can induce remissions in CLL patients with p53 abnormalities. Ann. Hematol. 82, 759–65.

    Article  PubMed  CAS  Google Scholar 

  89. Bosanquet, A.G. and Bell, P.B. (2004) Ex vivo therapeutic index by drug sensitivity assay using fresh human normal and tumor cells. J. Exp. Ther. Oncol. 4, 145–54.

    PubMed  CAS  Google Scholar 

  90. Tidefelt, U., Sundman-Engberg, B. and Paul, C. (1988) Effects of verapamil on uptake and in vitro toxicity of anthracyclines in human leukemic blast cells. Eur. J. Haematol. 40, 385–95.

    Article  PubMed  CAS  Google Scholar 

  91. Tidefelt, U., Sundman-Engberg, B., Rhedin, A.S. and Paul, C. (1989) In vitro drug testing in patients with acute leukemia with incubations mimicking in vivo intracellular drug concentrations. Eur. J. Haematol. 43, 374–84.

    Article  PubMed  CAS  Google Scholar 

  92. Lathan, B., von Tettau, M., Verpoort, K. and Diehl, V. (1990) Pretherapeutic drug testing in acute leukemias for prediction of individual prognosis. Haematol. Blood Transfus. 33, 295–8.

    PubMed  CAS  Google Scholar 

  93. Staib, P., Lathan, B., Schinköthe, T., Wiedenmann, S., Pantke, B., Dimski, T., Voliotis, D. and Diehl, V. (1999) Prognosis in adult AML is precisely predicted by the DISC-assay using the chemosensitivity-index Ci. Adv. Exp. Med. Biol. 457, 437–44.

    Article  PubMed  CAS  Google Scholar 

  94. Kirkpatrick, D.L., Chen, L. and Goh, T.S. (1992) Modification of the DiSC assay by the incorporation of monoclonal antibody staining. Leuk. Res. 16, 1097–103.

    Article  PubMed  CAS  Google Scholar 

  95. Beksac, M., Kansu, E., Kars, A., Ibrahimoglu, Z. and Firat, D. (1988) A rapid drug sensitivity assay for neoplasmatic cells. Med. Oncol. Tumor Pharmacother. 5, 253–7.

    PubMed  CAS  Google Scholar 

  96. Bosanquet AG, Raper, S.L, Durant, J., Scadding, S.M., Graham, D., Oscier, G.D., Richards, S.M., and Catovsky, D. (2006) Comparison of ex vivo drug sensitivity by TRAC assay and patient response in the UK LRF CLL4 trial. Haematologica/Hematol. J. 91(suppl 1), 100.

    Google Scholar 

  97. Bosanquet, A., Raper, S.L., Durant, J., Scadding, S.M., Graham, D., Oscier, G.D., Richards, S.M., and Catovsky, D. on Behalf of the NCRI CLL Working Group. (2006) Drug sensitivity by TRAC (DiSC) assay as a prognostic factor for patient response in untreated CLL: results from the UK LRF CLL4 trial. Blood 108, 94a.

    Google Scholar 

  98. Else, M., Smith, A.G., Raper, S.L., Kim S. and Cocks, S.C. and Catovsky, D. (2007) An association between drug sensitivity by TRAC (DiSC) assay and quality of life in the UK LRF CLL4 trial. 12th International Workshop on CLL, September 2007. www.caltri.org/pdf/ElseIWCLL07.pdf.

  99. Bosanquet, A.G., Richards, S.M., Wade, R., Else, M., Matutes, E., Dyer, M.J., Rassam, S.M., Durant, J., Scadding, S.M., Raper, S.L., Dearden, C.E. and Catovsky, D. (2009) Drug cross-resistance and therapy-induced resistance in chronic lymphocytic leukaemia by an enhanced method of individualised tumour response testing. Br. J. Haematol. 146, 384–95.

    Article  PubMed  CAS  Google Scholar 

  100. Staib, P., Staltmeier, E., Neurohr, K., Cornely, O., Reiser, M., Schinköthe, T. (2005) Prediction of individual response to chemotherapy in patients with acute myeloid leukaemia using the chemosensitivity index Ci. Br. J. Haematol. 128, 783–91.

    Article  PubMed  CAS  Google Scholar 

  101. Carstensen, H., and Tholander, B. (1985) Chemosensitivity of ovarian carcinoma: In vitro/in vivo correlations using the dye exclusion assay of Weisenthal. In: Proceedings: 3rd European Conference on Clinical Oncology. Stockholm, Sweden.

    Google Scholar 

  102. Nagourney, R.A., Brewer, C.A., Radecki, S., Kidder, W.A., Sommers, B.L., Evans, S.S., Minor, D.R. and DiSaia, P.J. (2003) Phase II trial of gemcitabine plus cisplatin repeating doublet therapy in previously treated, relapsed ovarian cancer patients. Gynecol. Oncol. 88, 35–9.

    Article  PubMed  CAS  Google Scholar 

  103. Carstensen, H. (1983) [Predictive testing of cytostatics in ovarian cancer using Weisenthal’s method]. Lakartidningen 80, 2812–6.

    PubMed  CAS  Google Scholar 

  104. Nagourney, R.A., Sommers, B.L., Harper, S.M., Radecki, S. and Evans, S.S. (2003) Ex vivo analysis of topotecan: advancing the application of laboratory-based clinical therapeutics. Br. J. Cancer 89, 1789–95.

    Article  PubMed  CAS  Google Scholar 

  105. Brewer, C.A., Blessing, J.A., Nagourney, R.A., Morgan, M. and Hanjani, P. (2006) Cisplatin plus gemcitabine in platinum-refractory ovarian or primary peritoneal cancer: a phase II study of the Gynecologic Oncology Group. Gynecol. Oncol. 103, 446–50.

    Article  PubMed  CAS  Google Scholar 

  106. Weisenthal LM, W.C., Smith ME, Sanchez CG, and Berglund RF. (2003) Platinum resistance determined by cell culture drug resistance testing (CCDRT) predicts for patient survival in ovarian cancer. http://weisenthal.org/w_ovarian_cp_toc.html.

  107. Pieters, R., Huismans, D.R., Loonen, A.H., Hählen, K., van der Does-van den Berg, A., van Wering, E.R. and Veerman, A.J. (1991) Relation of cellular drug resistance to long-term clinical outcome in childhood acute lymphoblastic leukaemia. Lancet 338, 399–403.

    Google Scholar 

  108. Weisenthal, L.M., Dill, P.L., and Pearson, F.C. (1990) Tumor and patient-specific activity of biologic response modifiers (ImuVert, tumor necrosis factor, alpha-interferon) in fresh specimens of human neoplasms detected by a sensitive and specific in vitro assay. Proc. Am. Assoc. Cancer Res. 31, 299.

    Google Scholar 

  109. Einhorn, S., Fernberg, J.O., Grandér, D. and Lewensohn, R. (1988) Interferon exerts a cytotoxic effect on primary human myeloma cells. Eur. J. Cancer Clin. Oncol. 24, 1505–10.

    Google Scholar 

  110. Lepri, E., Barzi, A., Menconi, E., Portuesi, M.G., Liberati, M. (1991) In vitro synergistic activity of PDN-IFN alpha and NM  +  IFN alpha combinations on fresh bone-marrow samples from multiple myeloma patients. Hematol. Oncol. 9, 79–86.

    Article  PubMed  CAS  Google Scholar 

  111. Windbichler, G.H., Hausmaninger, H., Stummvoll, W., Graf, A.H., Kainz, C., Lahodny, J., Denison, U., Müller-Holzner, E. and Marth, C. (2000) Interferon-gamma in the first-line therapy of ovarian cancer: a randomized phase III trial. Br. J. Cancer 82, 1138–44.

    Google Scholar 

  112. Alberts, D.S., Marth, C., Alvarez, R.D., Johnson, G., Bidzinski, M., Kardatzke, D.R., Bradford, W.Z., Loutit, J., Kirn, D.H., Clouser, M.C., Markman, M. for the GRACES Clinical Trial Consortium. (2008) Randomized phase 3 trial of interferon gamma-1b plus standard carboplatin/paclitaxel versus carboplatin/paclitaxel alone for first-line treatment of advanced ovarian and primary peritoneal carcinomas: results from a prospectively designed analysis of progression-free survival. Gynecol. Oncol. 109, 174–81.

    Article  Google Scholar 

  113. Weisenthal, L.M., Patel, N. and Rueff-Weisenthal, C. (2008) Cell culture detection of microvascular cell death in clinical specimens of human neoplasms and peripheral blood. J. Intern. Med. 264, 275–87.

    Article  PubMed  CAS  Google Scholar 

  114. Weisenthal, L., Lee, D.J., and Patel, N. (2008) Antivascular activity of lapatinib and bevacizumab in primary microcluster cultures of breast cancer and other human neoplasms, In: ASCO 2008 Breast Cancer Symposium. Washington, D.C. Abstract # 166.

    Google Scholar 

  115. Weisenthal, L. (2006) Gefitinib-induced cell death in short term fresh tumor cultures predicts for long term patient survival in previously-treated non-small cell lung cancer. J. Clin. Oncol.. 24(18S), 17117.

    Google Scholar 

  116. Selby, P., Buick, R.N. and Tannock, I. (1983) A critical appraisal of the “human tumor stem-cell assay”. N. Engl. J. Med. 308, 129–34.

    Article  PubMed  CAS  Google Scholar 

  117. Al-Nasiry, S., Geusens, N., Hanssens, M., Luyten, C. and Pijnenborg, R. (2007) The use of Alamar Blue assay for quantitative analysis of viability, migration and invasion of choriocarcinoma cells. Hum. Reprod. 22, 1304–9.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Weisenthal Cancer Group, Huntington Beach, CA, USA

    Larry M. Weisenthal

Authors
  1. Larry M. Weisenthal
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Larry M. Weisenthal .

Editor information

Editors and Affiliations

  1. Translation Oncology Research Centre, Queen Alexandra Hospital, Southwick Hill Road, Portsmouth, P06 3LY, United Kingdom

    Ian A. Cree

Rights and permissions

Reprints and permissions

Copyright information

© 2011 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Weisenthal, L.M. (2011). Differential Staining Cytotoxicity Assay: A Review. In: Cree, I. (eds) Cancer Cell Culture. Methods in Molecular Biology, vol 731. Humana Press. https://doi.org/10.1007/978-1-61779-080-5_22

Download citation

  • DOI: https://doi.org/10.1007/978-1-61779-080-5_22

  • Published:

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-079-9

  • Online ISBN: 978-1-61779-080-5

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation