A MINIATURIZED METHOD FOR THE KARYOTYPIC ANALYSIS OF BONE-MARROW OR BLOOD-SAMPLES IN HEMATOLOGICAL MALIGNANCIES

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.