A simple method for short-term culturing bone marrow and unstimulated blood from acute leukemias

It has become routine practice to culture bone marrow and/or unstimulated peripheral blood for cytogenetic analysis due to associations of consistent chromosomal abnormalities within specific subgroups of leukemias. Unfortunately, two leukemias for which numerous chromosomal aberrations have been recorded, acute myelogenous leukemia (AML) and acute lymphocytic leukemia (ALL), frequently present with very low cell counts often resulting in unsuccessful cultures. Furthermore, when cultures are successful, chromosomes are often contracted and metaphase spreads can be of poor quality. In the present investigation, by first treating blood or bone marrow from AML and ALL patients with ammonium chloride (1.5 mM) we were able to obtain chromosomal preparations often similar in quality to those obtained from normal stimulated blood cultures. With this improved culturing method it is also possible to generate high-quality chromosomes and DNA from the same sample.