A strategy combining the use of FIX&PERM® for the intracellular detection of the cell cycle marker Ki-67 and 10 colour flow cytometry of cell surface markers was used to determine reference values for the proliferative activity of hematopoietic cells in normal bone marrow.
Hematologic malignancies such as leukaemia and lymphomas are forms of cancer that begin in the cells of blood-forming tissues like the bone marrow. Ki-67 is a cell proliferation marker located in the nucleus, and expression can be indicative of clinical outcome in patients with these blood-borne cancers. Ki-67 detection is most commonly performed in histopathology using an immunohistochemical approach, while use in flow cytometry has historically been sporadic due to the limited number of parameters that could be detected in the heterogeneous bone marrow cell population.
The aim of the flow cytometry study by Nies, et al. was that in order to understand Ki-67 expression using flow cytometry analysis in cancer patients, it is first necessary to determine the reference values for healthy individuals for the different hematopoietic cell lineages. This includes Ki-67 expression during the cell maturation stages of myelopoiesis, monopoiesis, and erythropoiesis. Healthy bone marrow was taken from the femur of hip replacement patients and a strategy of gating the different cell populations was based on cell surface markers. FIX&PERM was used in combination with Ki-67 immunostaining and flow cytometry software to determine the proliferation changes during the different maturation pathways. As a result it was possible to determine reference values for this cell cycle parameter in the different blood cell lineages and stages of maturation.
For data and methods see: Nies, K.P.H. et al. Determination of the proliferative fractions in differentiating hematopoietic cell lineages of normal bone marrow. Cytometry A. 2018 93(11):1097-1105. PMID: 30176186.
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