Dna Synthesis Time in Leukaemic Cells as Measured by the Double Labelling and the Percentage Labelled Mitoses Methods

Harriss, E. B.; Hoelzer, Dieter

doi:10.1111/j.1365-2184.1971.tb01552.x

Cited by 15 publications

(6 citation statements)

References 14 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…injection of leukaemic cells. The rate of develop ment is closely correlated with the number of cells transferred, and it is very reproducible in its course and its kinetics [9,10]. Death of leukaemic rats occurs with a leukocyte count of about 400,000 cells/mm3.…”

Section: Methodsmentioning

confidence: 88%

The Failure of Normal Haemopoiesis in Rats during the Development of Acute Leukaemia

Hoelzer¹,

Harriss²

1973

Acta Haematol

Self Cite

View full text Add to dashboard Cite

The reaction of normal haemopoiesis under the development of a transferable acute leukaemia was studied in the rat. A reduction of cell numbers was observed in the bone marrow in the proliferating compartments of erythropoiesis, megakaryocytopoiesis and myelopoiesis. In the peripheral blood, erythrocyte and platelet levels decreased to one tenth of normal: Granulocytes and lymphocytes showed an increase coincident with the increase in circulating leukaemic cells before a final tendency to decline. These changes are attributed partly to variations in the inflow from the bone marrow and partly to direct effects on the peripheral blood cells themselves. The reduced number of cells in the proliferation compartment may be brought about by effects on this compartment itself, but there are some indications that a reduced inflow from the preceding stem cell compartment might be responsible.

show abstract

Section: Methodsmentioning

confidence: 88%

The Failure of Normal Haemopoiesis in Rats during the Development of Acute Leukaemia

Hoelzer¹,

Harriss²

1973

Acta Haematol

Self Cite

View full text Add to dashboard Cite

show abstract

“…The double-label studies described in the literature to study cell kinetics (6,7,18,20) involved either 14Cor 3Hthymidine infusions. Because only very small amounts of radioisotopes could be administered to patients, radioautographs often had to be left in the dark for several months.…”

Section: Discussionmentioning

confidence: 99%

Double labeling and in vitro versus in vivo incorporation of bromodeoxyuridine in patients with acute nonlymphocytic leukemia

1985

View full text Add to dashboard Cite

A monoclonal antibody against bromodeoxyuridine (BrdUrd) was produced, and a rapid slide technique (RPMB technique) was developed for the estimation of S-phase cells in a population using this antibody. Bone marrow cells from patients with acute nonlymphocytic leukemia (ANLL) were studied by both the RPMB technique and tritiated thymidine (3HdThd) labeling index studies. The percentage of S-phase cells obtained by each method was compared in 50 samples, and the correlation coefficient was r = 0.89.A "double label" method is also described in which cells were simultaneously incubated with either BrdUrd and 3HdThd or BrdUrd and tritiated cytosine arabinoside (3HAra-C).The samples were first processed by the Key terms: Anti-BrdUrd antibody, double labeling, acute nonlymphocytic leukemia, cell cycle characteristics, cytosine arabinoside Certain halogenated pyrimidines such as 5-bromo-2'deoxyuridine (BrdUrd) and 5-iodo-2'-deoxyuridine (IUdR) can be incorporated into DNA in place of thymidine, and consequently they have been used for studies of DNA replication for a number of years (1,11,17). Both polyclonal (4,16) and monoclonal (3,12) antibodies have been developed against these compounds and have been successfully utilized for quantitating the proportion of S-phase cells in a population (5) and for studying effects of drugs on DNA synthesis of cells (2) and the frequency of sister chromatid exchanges (9).We developed a rapid slide technique (RPMB technique) for determination of S-phase cells in a population using a monoclonal anti-BrdUrd antibody (RPMB I). In this immunofluorescent assay, every S-phase cell demonstrated bright green fluorescence. Additionally, we were able to quantitate fluorescence per cell by using an automated microphotometer system attached to the fluorescence microscope, and we showed that the degree of fluorescence was directly proportional to the amount of DNA synthesis in individual cells.A "double-label" technique was then established by incubating freshly obtained human leukemic cells with BrdUrd and tritiated thymidine (3HdThd) simultaneously. The samples were first processed by RPMB technique, so that every S-phase cell revealed bright green fluorescence. Following the application of photographic emulsion and development by autoradiography (ARG),we were able to demonstrate black grains of 3HTdR overlying the nucleus of every fluorescent cell. This double label technique not only validated the RPMB technique but allowed us to introduce two probes simultaneously into the DNA.

show abstract

“…The administration of the first label in uiuo, followed by a second label in vitro, is probably less liable to error, since the progression of cells is then permitted to occur in the animal (e.g. Harris & Hoelzer, 1971).…”

Section: Discussionmentioning

confidence: 99%

IN VITRO AND IN VIVO LABELLING OF ANIMAL TUMOURS WITH TRITIATED THYMIDINE

Denekamp

Kallman

1973

Cell Proliferation

View full text Add to dashboard Cite

The labelling indices obtained by incubating tumour specimens with tritiated thymidine in vitro under hyperbaric oxygen have been compared with those obtained by labelling a matched tumour in vivo. The correspondence between these individual labelling indices is sometimes very poor; however, the average labelling index derived from groups of tumours labelled in the two ways does not differ significantly. There was a large variation from field to field within any tumour, and considerable variation from one tumour to another within each tumour type. The mitotic index was also compared in the matched tumour preparations; the mitotic index in vitro was almost always considerably lower than the values observed in vivo.

show abstract

Dna Synthesis Time in Leukaemic Cells as Measured by the Double Labelling and the Percentage Labelled Mitoses Methods

Cited by 15 publications

References 14 publications

The Failure of Normal Haemopoiesis in Rats during the Development of Acute Leukaemia

The Failure of Normal Haemopoiesis in Rats during the Development of Acute Leukaemia

Double labeling and in vitro versus in vivo incorporation of bromodeoxyuridine in patients with acute nonlymphocytic leukemia

IN VITRO AND IN VIVO LABELLING OF ANIMAL TUMOURS WITH TRITIATED THYMIDINE

Contact Info

Product

Resources

About