The gubernaculum testis is the connective tissue organ that causes the testis to descend. How the process of testicular descent is regulated is not fully understood. Current hypotheses postulate that a nonandrogenic fetal testicular factor controls the first phase of descent, that is characterized by growth of the gubernaculum and transabdominal migration of the testis. When gonadal extracts from fetuses with ages corresponding to the first phase of testicular descent (50, 60, and 75 days) were tested on gubernacular cells, the growth stimulatory effect of testicular extracts exceeded the effect of both ovarian extract and fetal calf serum. Gonadal extracts from 80-, 90-, and 100-day-old fetuses showed only a minor sex difference. No sex difference or age-dependent changes were detected when fetal gonadal extracts were tested on murine 3T3 cells. Polypeptide growth factors (epidermal growth factor, insulin, fibroblast growth factor, platelet-derived growth factor, and transforming growth factor-beta) were tested for growth stimulatory activity and had only minor effects on gubernaculum cells. Fetal testicular hormones (anti-Müllerian hormone, inhibin, and androgenic steroids) did not induce initiation of DNA synthesis at concentrations that are highly bioactive in typical target systems. When testicular samples were dialyzed, the high mol wt fraction (greater than 3500) had lower growth stimulatory activity in gubernaculum cells, but not 3T3 cells. Bioactivity of ovarian extracts and fetal calf serum was not diminished after dialysis. The low mol wt fraction (less than 3500) of testicular extract was distinctly stimulatory to gubernaculum cells but not 3T3 cells, and the low mol wt fraction of ovarian extracts did not stimulate growth in either cell type. It was concluded that the fetal porcine testis during the first phase of testicular descent contains low mol wt factor(s) to which gubernaculum cells and not 3T3 cells are responsive. The bioactive fraction probably contains the factor(s) that initiate testicular descent. We suggest the name descendin for this new activity.