The use of peripheral blood feeder layers as a source of GM-CSF for human bone marrow cultures

The use of peripheral blood feeder layers as a source of stimulus for colony formation by human granulocytic progenitor cells in semi-solid agar cell cultures was examined. Comparison with various conditioned media demonstrated that cultures stimulated by peripheral blood feeder layers produced the greatest number and largest colonies. The cell concentration in the feeder layers was more important than the total cell number. Feeder layer plates containing 2 X 10(5) cells at a concentration of 1 X 10(6) cells/ml proved to be just potent as the conventional feeder layer plates containing 1 x 10(6) cells/ml in 1 ml. Thus feeder layer plates can be more economical in terms of cell numbers than has previously been reported. By using a known panel of donors for the leukocytes, the number of sub-optimal batches of feeder layers was reduced. Addition of 1 microM adenosine 3':5'-cyclic-monophosphate or 10 micrograms/ml Li2CO3 to poor feeder layers enhanced their colony-stimulating ability.