Measurement of CD34+ cells in bone marrow by flow cytometry.
A procedure is described for the measurement of the %CD34+ progenitor cells in bone marrow using directly conjugated antibodies. Staining cells with anti-CD45.FITC in conjunction with anti-CD34.PE allows the CD45- nucleated red blood cells and the CD45++ lymphocytes and monocytes to be separated from the CD45+ progenitor cells. Granulocytes are separated from the CD34+ cells based on differences in side scatter properties. A gated acquisition of CD34+ cells is used to define the boundaries of the CD34+ population in a plot of forward scatter vs side scatter and in a plot of anti-CD45.FITC vs anti-CD34.PE. Use of these regions during analysis reduces background staining and allows for a consistent identification of a CD34+ population. Acquisition of 50,000 cells provides adequate precision of the %CD34+ measurement. Acquisition and analysis procedures are presented for use of both a Becton Dickinson FACScan flow cytometer and a Coulter EPICS Profile II flow cytometer.
Trischmann
T M
TM
Johns Hopkins Oncology Center, Baltimore, MD 21287.
Schepers
K G
KG
Civin
C I
CI
D000911:Antibodies, Monoclonal / Q000276:immunology; D015703:Antigens, CD / Q000032:analysis* / Q000276:immunology; D018952:Antigens, CD34; D001854:Bone Marrow Cells; D002452:Cell Count / Q000379:methods*; D002469:Cell Separation; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate; D005455:Fluorescent Antibody Technique; D006412:Hematopoietic Stem Cells / Q000737:chemistry; D006801:Humans; D017493:Leukocyte Common Antigens / Q000032:analysis; D012984:Software
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D018952:Antigens, CD34; D017493:Leukocyte Common Antigens; D016650:Fluorescein-5-isothiocyanate