Growth of erythroid colonies in agar cultures of normal human bone marrow

The use of methylcellulose (MC) gels or plasma clots, for the support of human erythropoiesis in vitro, is associated with several technical disadvantages. Substitution of soft agar offers the prospect of overcoming these difficulties. In comparative studies, normal human bone marrow cells were cultured with erythropoietin (Epo) in agar (0.1%-0.3%) and MC. Concentrations of 0.175% and 0.2% agar proved to be optimal with respect to the combination of cloning efficiency and colony density. Further morphological examination revealed that subcolony formation in erythroid 'bursts' was influenced by gel viscosity. In additional experiments, miniaturising the assay system, to 0.25 ml culture volumes, increased cloning efficiency and reduced Epo utilization. These results confirm and expand earlier observations, and support a preference for the general use of agar in human erythroid cell cultures.