SUMMARY The entire second thoracic mammary glands of 4-week-old BALB/c female mice primed with oestradiol plus progesterone were cultivated in organ culture medium containing the 'growth-promoting' hormone combinations: insulin, prolactin, growth hormone, oestradiol, progesterone and aldosterone or insulin, prolactin and aldosterone. Full lobulo-alveolar development was induced after 5–6 days of incubation and could be maintained for 15–16 days in organ culture in medium containing either

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... e combination. After the initial 5–6 days in the 'growth-promoting' medium, subsequent cultivation of the glands in a medium with the 'lactogenic hormones', insulin, prolactin plus cortisol, led to accumulation of 'milk-like' secretory material in the ductal and alveolar lumina. Incubation of the lobulo-alveolar gland in medium with insulin alone for 7–9 days resulted in complete regression of the alveoli leaving only a ductal parenchyma. Incubation in insulin, prolactin, growth hormone or insulin plus the steroid hormones for 7–9 days led to considerable alveolar degeneration without a complete regression. The results indicate that both pituitary and steroid hormones are essential for development and maintenance of mammary alveoli; insulin can only sustain the basal ductal structure.