Colony Formation in Culture by Bovine Granulopoietic Progenitor Cells

Calf bone marrow cells cultured in a semi-solid medium of 0.8% methyl cellulose produced colonies of granulocytic cells and macrophages by seven days. A prerequisite for colony growth was the presence of serum obtained from a calf three hours after intravenous injection of endotoxin. Three morphological types of colonies were seen but cell types within these types of colonies did not differ. Cultured cells were identified by morphological and cytochemical characteristics.

Optimum growth occurred when serum from endotoxin stimulated calves and fetal calf serum were present in a volumetric ratio of 7:3. Inhibition of colony growth occurred when endotoxin-stimulated serum was present at greater than optimum concentration. Normal calf serum, fetal calf serum, mouse L-cell conditioned medium and bovine urine did not stimulate significant colony growth when 8.0 x 10⁴ marrow cells were cultured.

There was a linear relationhip between the number of marrow cells in the cultures and the number of colonies produced. Colony forming efficiency ranged from 13 to 59 colonies per 10⁵ cells plated.

The behaviour of calf colony forming units in suspension culture was similar to that reported for mouse colony forming units.