Shahida Mohsin, Wim Renmans, Serge Damiaens, Ivan Van Riet, Rik Schots, Marc De Waele.
Enumeration of haematopoietic progenitor cells with flow Cytometry: Comparison of different methods.
Pak J Pathol Jan ;10(2):16-5.

In this study several flow cytometric methods to enumerate peripheral blood hematopoietic progenitor cells were compared. Blood samples taken during stem cell mobilisation in eight patients with haematological or solid neoplasms and in one allogeneic donor were examined. Whole blood and mononuclear cell suspensions were labelled with FITC and PE conjugates of the HPCA2 (CD34) monoclonal antibody. The percentage of CD34 positive cells was determined in all nucleated cells and in an enlarged lymphocyte gate. Absolute numbers were calculated. A good correlation was found between the results of the different methods. A CD34-PE conjugate gave higher fluorescence intensity than CD34-FITC but induced more background staining. An enlarged lymphocyte gate could be defined which included all CD34 cells. Absolute numbers could then be calculated by taking into account the number of monocytes in the gate. Ficoll-Hypaque centrifugation did not induce selective loss of precursors. Correction for red blood cell contamination based on the results of CD45 staining, was of minor clinical importance in these samples. In conclusion, a whole blood lysis method with a CD34-FITC conjugate and analysis of all nucleated cells can be recommended.

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