Fibroblasts isolated by enzymic digestion of chick embryo tendons were incubated for several hours in suspension under conditions in which they were in steady state in terms of the synthesis and secretion of procollagen. Under these conditions, the cells synthesized and secreted about 630 microgram of procollagen/10(9) cells/h. The cells were labeled with [14C]proline for 15 to 120 min and then the kinetics of secretion were followed by chasing the label and assaying the 14C-peptides digestible by collagenase in the cells and in the medium. The results demonstrated that secretion of collagenase-digestible peptides did not follow the kinetics of a single first order process but suggested at least two pseudo-first order process with half-times of 14 and 115 min. The [14C]procollagen secreted during 0 to 30 min and 90 to 120 min of chase was the same in terms of the ratio of pro-alpha1 to pro-alpha2 chains, the size of the pro-alpha chains, the extent of interchain disulfide bonding, the extent of prolyl hydroxylation, and the degree of helicity as tested by resistance to pepsin digestion. Addition of ascorbic acid to the incubation medium increased slightly the extent of prolyl hydroxylation but did not alter the kinetics of secretion. The results suggested that the kinetics of secretion are influenced by a two-compartment system in which at least one metabolic pool contributing to the secretory process is present as a "side pocket."