Enrichment of murine granulomonocytic progenitors using a continuous linear Ficoll-Isopaque gradient

Murine bone marrow was fractionated using a Ficoll-Isopaque continuous linear gradient characterized by an osmolarity of 290 mOsm constant throughout the gradient and a physiological pH of 7.4. The cellular peak detected prior to fractionation by means of a 405 nm light beam served as a guide for determining fraction collection. Under these conditions CFC enrichment was observed constantly for densities lower than that of the cellular peak. In 17 experiments the enrichment factor amounted to 3.6 +/- 1.4. Enrichment appeared to be due to both an increase in CFC concentration and an improvement in CFC cloning efficiency. A correlation between the concentration of CFC and that of undifferentiated blasts was observed. The CFC density distribution was dependent on the cell load. For cell loads lower than 25 x 10(6) the modal density of CFC was within the range 1.0615-1.0715. For cell loads higher than 25 x 10(6) there was a shift of the distribution curve towards higher density. Clu-CFC appeared to be denser than CFC. This increase in density may be due to a maturation process from CFC to Clu-FC as a maturation gradient with increasing density was found for the granulocytic series and for erythroblasts.