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Laboratoire de Physiologie de la Lactation, Institut National de la Recherche Agronomique, C.N.R.Z., 78350 Jouy-en-Josas, France

(Received 8 June 1976)

Previous studies have established that in the presence of insulin, mouse and rat mammary cells synthesize DNA and undergo mitosis in vitro. In cultured mammary tissue from virgin mice this event is critical for the subsequent appearance of secretory activity in daughter cells in response to adrenal corticosteroids and lactogenic hormones (Juergens, Stockdale, Topper & Elias, 1965; Stockdale & Topper, 1966; Turkington, 1968), whereas in cultures from pregnant mice the critical mitosis has already occurred in vivo (Owens, Vonderhaar & Topper, 1973). Mammary gland explants from 12-day pseudopregnant rabbits (Delouis & Denamur, 1972) are well maintained in Medium 199 (Microbiological Associates) in the presence of insulin in atmospheres containing 20, 57 or 95% O₂ and the addition of prolactin stimulates secretory activity under these conditions (Delouis, 1974). In the