Cell kinetics with in vivo bromodeoxyuridine and flow cytometry: Clinical significance in acute non-lymphoblastic leukaemia

Riccardi, Alberto; Giordano, Monica; Danova, Marco; Girino, Margherita; Brugnatelli, Silvia; Ucci, G.; Mazzini, Giuliano

doi:10.1016/0277-5379(91)90139-5

Cited by 24 publications

(14 citation statements)

References 21 publications

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“…Although there are several clinical studies published using this technique on a variety of tumour types (Wilson et al, 1988;Begg et al, 1990;Rew et al, 1991;Riccardi et al, 1991;Rew et al, 1992), the reliability and reproducibility of the method has yet to be examined in detail. It has been demonstrated that the measurement of ploidy may be performed accurately both between and within institutions (Wheeless et al, 1991).…”

Section: Discussionmentioning

confidence: 99%

An assessment of the reliability and reproducibility of measurement of potential doubling times (Tpot) in human colorectal cancers

Wilson

West²,

Wilson³

et al. 1993

Br J Cancer

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Summary An assessment has been made of the reproducibility of measuring tumour proliferation using in vivo iododeoxyuridine (IUdR) dissolved in 10 ml of sterile water. This was performed with informed consent and Hospital Ethical Committee approval. The tumours were then either biopsied (n = 28) or resected (n = 25) after a mean interval of 6.2 h (range 3.0-14.7 h) following injection. This interval (t) was measured from the time of IUdR administration as a bolus over approximately 30s until the cessation of the sample's blood supply. The samples were then immediately fixed in 70% ethanol and stored at 4°C. Parallel samples taken from adjacent tissue were fixed in formol saline for pathological assessment. No patient had received radio-or chemotherapy prior to IUdR labelling and tumour sampling.On the day of analysis, samples were digested to produce a nuclear suspension and stained for IUdR and total DNA content as previously described (Wilson et al., 1985). In summary, the specimens were minced, then digested using 0.4 mg ml' pepsin in 0.1 M HCl and the DNA partially denatured using 2 M HCI for 15 min to expose IUdRincorporated DNA. Approximately 2 x 106 nuclei were then incubated with an anti-IUdR monoclonal antibody (BecktonDickinson) for 1 h at room temperature (1 in 20 dilution). After washing, the nuclei were incubated with a rabbit antimouse IgG FITC conjugate (Dakopatts) for a further 30 min (1 in 40 dilution). Finally, the nuclei were counterstained with propidium iodide at a concentration of 10 gmlm' (Sigma) to allow measurement of the total DNA content.The method of data analysis has previously been described (Wilson et al., 1985). The analyses were performed at the Gray Laboratory, Northwood on an Ortho Systems 50-H Cytofluorograph (Beckton-Dickinson) and at the Paterson Institute for Cancer Research, Manchester, using an EPICS V flow cytometer (Coulter). Both machines created light with a wavelength of 488 nm from 5 W argon ion lasers operating at 200 mW; green fluorescent light emitted by excitation of the FITC conjugate bound to the anti-IUdR monoclonal antibody was collected at 510-560 nm and red fluorescence from the excited PI above 620 nm. All data were collected from the cytometers on 1024 channels using list mode. The data analysis requires the use of software which allows regions (gates) to be set around various populations of particles. This can be used to remove debris or clumps of nuclear material from the analyses to facilitate estimations of the proportions of nuclei within each part of the cell cycle and their mean DNA content. The data were gated to exclude multiple nuclei and debris on the DNA peak vs area signals.

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Section: Discussionmentioning

confidence: 99%

An assessment of the reliability and reproducibility of measurement of potential doubling times (Tpot) in human colorectal cancers

Wilson

West²,

Wilson³

et al. 1993

Br J Cancer

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“…The ability to accurately detect patients who may need and respond favourably to additional treatment would be an advantage and would increase the likelihood of determining potential benefits of adjuvant therapy. The measurement of tumour proliferation may help in the identification of tumours which, because of their rapidly proliferative nature, might benefit from modification of their radiotherapy (Dische & Saunder, 1989); and secondly may be more susceptible to chemotherapy than less proliferative lesions (Riccardi et al, 1991).…”

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confidence: 99%

Intra-tumoral heterogeneity of tumour potential doubling times (Tpot) in colorectal cancer

Wilson

West²,

Wilson³

et al. 1993

Br J Cancer

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“…Murovic et al (1986) found that paediatric medulloblastomas, glioblastomas multiforme and highly anaplastic astrocytomas had high proliferative potentials compared with the adult forms of the same tumours. In a more recent study Danova et al (1991) compared DNA index, LI, Tpo t and histology in 68 cases of astrocytomas and found that mean survival time was correlated with these parameters. (1985,1988) found that various CNS tumours, for example ependymomas, mixed gliomas and moderately anaplastic astrocytomas, exhibited very low LIs (i.e.…”

Section: Tissue Sites Of Specific Tumoursmentioning

confidence: 99%

“…S. Nakamura et al (1991) used in vivo BrdUrd labelling and double bone marrow aspirations to determine Ts (9.7 h) and T c (152.5 h) in bone marrow cells of untreated patients with acute leukaemia. Riccardi et al (1986Riccardi et al ( , 1988Riccardi et al ( , 1989Riccardi et al ( , 1991 applied flow cytometric methods to the kinetic analysis of acute leukaemias, AML and ANLL. An additional observation reported in the same study was that of labelled granulocytes, indicating either possible in vivo differentiation of leukaemic precursors or normal haematopoietic precursors.…”

Section: Hematopoietic Tumoursmentioning

confidence: 99%

Bromodeoxyuridine: a diagnostic tool in biology and medicine, Part II: Oncology, chemotherapy and carcinogenesis

Dolbeare

1995

Histochem J

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Cell kinetics with in vivo bromodeoxyuridine and flow cytometry: Clinical significance in acute non-lymphoblastic leukaemia

Cited by 24 publications

References 21 publications

An assessment of the reliability and reproducibility of measurement of potential doubling times (Tpot) in human colorectal cancers

An assessment of the reliability and reproducibility of measurement of potential doubling times (Tpot) in human colorectal cancers

Intra-tumoral heterogeneity of tumour potential doubling times (Tpot) in colorectal cancer

Bromodeoxyuridine: a diagnostic tool in biology and medicine, Part II: Oncology, chemotherapy and carcinogenesis

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