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Fig. 1. MIAMI cells. (A) The number of colonies obtained after density gradient centrifugation (black bars) was significantly smaller than the number of colonies obtained without centrifugation (white bars) after plating the same number of bone marrow cells per dish using the conditions for expansion/selection of MIAMI cells. This result was observed in all cultures examined independent of age. The graph shows the mean±s.e.m. of more than 15 representative experiments. (B) MIAMI cells are enriched by a unique expansion/selection procedure to isolate a population of small (7-10 µm), highly proliferative cells with reduced cytoplasm. Essential in this procedure is a unique growth environment, with sequential culturing steps under defined oxygen tension, cell density, growth factors, serum concentration and substratum conditions. (C) MIAMI cells plated at low density (0.1-0.2 cells/cm2) were allowed to proliferate (D) and when single-cell-derived colonies reached a size of 50-100 cells (E) the colonies where isolated using cloning rings, replated and expanded again at low density. The morphology of the cells is maintained after 5 cell doublings (F) and 52 cell doublings (G). (B,F,G) 20x magnification; (C-E) 10x magnification.





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