C. Brigaudeau et al., A MINIATURIZED METHOD FOR THE KARYOTYPIC ANALYSIS OF BONE-MARROW OR BLOOD-SAMPLES IN HEMATOLOGICAL MALIGNANCIES, HEM CELL TH, 38(3), 1996, pp. 275-277
Standard techniques of karyotypic analysis of bone marrow or periphera
l blood cells generally use large numbers of cells. Thus, the quantity
of cells harvested from one bone marrow or blood puncture frequently
represents a limiting factor for other assays such as molecular biolog
y or flow cytometry, which are often essential for the diagnosis of he
matological diseases. To resolve this problem, we developed a ''miniat
urized technique'' of cytogenetic analysis that we tested on bone marr
ow (BM) cells from 20 patients with multiple myeloma (MM), 5 patients
with acute leukemia (AL), as well as on CD34+ cells purified from the
blood of 8 patients with agnogenic myeloid metaplasia (AMM). We used 3
x 10(6) BM cells from MM patients (for testing 6 different culture co
nditions), 10(6) BM cells from AL (2 culture conditions) and 4 x 10(3)
CD34+ cells from AMM patients. In most patients, 20 good quality meta
phases per slide were easily analyzed, showing that a 10-40 times redu
ction of the number of cells used for cytogenetics allows a reliable k
aryotypic analysis in hematological malignancies.