The influence of sample source and cell concentration on the in vitro chemosensitivity of haematological tumours

Bird, Martin C.; Forskitt, Susan; Gilby, Edward D.; Bosanquet, AG

doi:10.1038/bjc.1986.84

Cited by 8 publications

(3 citation statements)

References 12 publications

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“…The correlation for all 106 paired BM and PB LC50 comparisons was very good, with most pairs close to the ideal line x = y (Figure 3). Our findings agree with those of Bird et al (1986), who reported a significant association in sensitivity to a maximum of six drugs of leukaemic BM and PB cells from 12 patients. The same was found by Sargent and Taylor (1989) and Spiro et al (1981a), in single cases.…”

Section: Reagentssupporting

confidence: 80%

In vitro drug sensitivity of normal peripheral blood lymphocytes and childhood leukaemic cells from bone marrow and peripheral blood

Kaspers

Pieters²,

Zantwijk³

et al. 1991

Br J Cancer

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Summary In vitro drug sensitivity of leukaemic cells might be influenced by the contamination of such a sample with non-malignant cells and the sample source. To study this, sensitivity of normal peripheral blood (PB) lymphocytes to a number of cytostatic drugs was assessed with the MTT assay. We compared this sensitivity with the drug sensitivity of leukaemic cells of 38 children with acute lymphoblastic leukaemia. We also studied a possible differential sensitivity of leukaemic cells from bone marrow (BM) and PB. The following drugs were used: Prednisolone, dexamethasone, 6-mercaptopurine, 6-thioguanine, cytosine arabinoside, vincristine, vindesine, daunorubicin, doxorubicin, mafosfamide (Maf), 4-hydroperoxy-ifosfamide, teniposide, mitoxantrone, L-asparaginase, methotrexate and mustine.Normal PB lymphocytes were significantly more resistant to all drugs tested, except to Maf. Leukaemic BM and PB cells from 38 patients (unpaired samples) showed no significant differences in sensitivity to any of the drugs. Moreover, in 11 of 12 children with acute leukaemia of whom we investigated simultaneously obtained BM and PB (paired samples), their leukaemic BM and PB cells showed comparable drug sensitivity profiles. In one patient the BM cells were more sensitive to most drugs than those from the PB, but the actual differences in sensitivity were small.We conclude that the contamination of a leukaemic sample with normal PB lymphocytes will influence the results of the MTT assay. The source of the leukaemic sample, BM or PB, does not significantly influence the assay results.

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Section: Reagentssupporting

confidence: 80%

In vitro drug sensitivity of normal peripheral blood lymphocytes and childhood leukaemic cells from bone marrow and peripheral blood

Kaspers

Pieters²,

Zantwijk³

et al. 1991

Br J Cancer

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“…Leukaemic blasts from bone marrow and peripheral blood also behave similarly in the DiSC assay (Bird et al, 1986).…”

Section: Appraisal Of Methodsmentioning

confidence: 95%

Appraisal of the MTT assay as a rapid test of chemosensitivity in acute myeloid leukaemia

Sargent¹,

Taylor²

1989

Br J Cancer

293

220

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Summary We describe the application of a simple, rapid, semi-automated assay to the sensitivity testing of cytotoxic drugs in 23 patients with acute myeloid leukaemia (AML). The survival of blast cells from the bone marrow was measured by the MTT assay after 48h continuous exposure to drugs both singly and in combination. There was a linear relationship between the number of leukaemic ceils and the optical density of the Cytotoxic drug therapy remains the prime method of treatment in acute myeloid leukaemia. One of the recognised limitations of this therapy, however, is the inability to predict tumour sensitivity in individual patients. Most attention in the field of haematological malignancies has focused on the clonogenic assay (Marie et al., 1987), dye exclusion assays (Weisenthal et al., 1986;Bird et al., 1988) and radioactive precursor incorporation (Schwarzmeier et al., 1984;Raza et al., 1987). The advantages and disadvantages of these methods are well documented (Hill, 1983; Wiesenthal & Lippman, 1985). A simple rapid chemosensitivity test suitable for automation is what is required for routine use. The clonogenic assay is long-term, effectively measuring the chemosensitivity of dividing cells only. Evidence is emerging to suggest that non-clonogenic assays which measure cell kill in the total blast cell population may be equally valuable. The most promising of these, the dye exclusion assay, shows a good correlation with the end-point of the clonogenic assay (Weisenthal et al., 1983). However, it is not an automated technique and is therefore timeconsuming and subject to observer error.In 1983 Mosmann described a semi-automated colorometric assay based on the premise that the mitochondria of living cells reduce the tetrazolium salt MTT to formazan. A modified form of this is currently being successfully applied by the National Cancer Institute USA to the chemosensitivity testing of new drugs on cell lines (Alley et al., 1988). The technique has been adapted for chemosensitivity testing of chronic (Twentyman et al., 1989) and acute (Pieters et al., 1988) lymphatic leukaemia cells. Results compared favourably to those using the differential staining cytotoxicity (DiSC) assay, a dye exclusion technique (Pieters et al., 1989).It is important to validate this assay for each cell type, and consequently we describe its application to the chemosensitivity testing of blast cells from the bone marrow of patients with acute myeloid leukaemia. These patients have a poor prognosis even in those who achieve remission. A simple in vitro method aiding initial selection of drugs both singly and in combination and permitting retesting throughout remission induction and on relapse would be a therapeutic advance. Thirteen patients were assayed both on presentation and throughout the course of the disease, two after remission was already established, three after relapse and the remaining five patients tested initially had no follow-up as they did not survive beyond the first course of treatment. Methods Preparation of ce...

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“…We have had good success whether measured by the proportion of tests that give successful results (up to 95%; Bird et al, 1986a) or the correlation of assay results with patient response (overall 86% true positive and true negative correlations; Bosanquet et al, 1983;Bird et al, 1985Bird et al, , 1986bBird et al, , 1988.…”

Section: Introductionmentioning

confidence: 99%

Effect of cell isolation methods and drug concentration on the use of the Differential Staining Cytotoxicity (DiSC) assay with solid tumours

Bosanquet

Forskitt

1989

Cytotechnology

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The Differential Staining Cytotoxicity (DiSC) in vitro drug sensitivity assay has been tested in solid tumours obtained from surgery. Various enzyme cocktails have been used for disaggregating cells for times ranging from 1-72 h. Collagenase (0.5%) plus DNase (0.002%) was chosen as producing the best results when incubated for 3 h at 37°C. This cocktail was also used for 24, 48 and 72 h incubations. Cell suspensions so produced were sometimes cleaned up by centrifugation on a Nycodenz gradient. Breast tumours were more often successfully cultured (67%) compared to gastrointestinal (43%) or other tumours (34%). Results with 6 drugs tested in 10 or more tumours suggested that in general, higher drug concentrations should be used when testing solid tumours as opposed to leukaemias in the DiSC assay.

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The influence of sample source and cell concentration on the in vitro chemosensitivity of haematological tumours

Cited by 8 publications

References 12 publications

In vitro drug sensitivity of normal peripheral blood lymphocytes and childhood leukaemic cells from bone marrow and peripheral blood

In vitro drug sensitivity of normal peripheral blood lymphocytes and childhood leukaemic cells from bone marrow and peripheral blood

Appraisal of the MTT assay as a rapid test of chemosensitivity in acute myeloid leukaemia

Effect of cell isolation methods and drug concentration on the use of the Differential Staining Cytotoxicity (DiSC) assay with solid tumours

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