Chromosome preparation for myeloid malignancies

Abstract

Many cases of myeloid malignancies are associated with recurring cytogenetic aberrations. Chromosomal analysis can aid in diagnosis, predict prognosis, and disclose subsequent clonal evolution. Three different cell culture methods: direct harvest, nonsynchronized culture, and synchronized culture are usually prepared if the nucleated cell counts in marrow blood are sufficient. Synchronized culture is the first choice of method in myeloid malignancies, whereas the direct method can be omitted if the cell count is low. The aseptic culture technique is strictly followed until harvesting procedure. For synchronized culture, uridine and fluorodeoxyuridine are added as blocking reagents and released by thymidine on the following day. Harvesting steps of the cultures involved colcemid exposure, hypotonic treatment, and Carnoy’s fixation. The cells are then ready for slide making and banding for chromosomal analysis.