Bone marrow collection

In general, mononuclear cells (MNC) and metastatic cancer cells are separated from the bone marrow samples using a density-gradient like Ficoll or Lymphoprep. After centrifugation, the interface cells are removed and washed. For ICC analyses, known quantities of cells are deposited onto glass slides by cytocentrifugation. Preparing cytospins has the advantages that only a single layer of cells is present which allows quantification of tumour cells in relation to the number of MNC. The cell pellets can also be resuspended in RNA stabilising buffers for molecular down stream applications.