Differentiation of mouse mammary gland in vitro requires insulin, hydrocortisone, and prolactin. The epithelial cells must first divide in order to synthesize casein in response to these hormones. Insulin is required for the initiation of DNA synthesis and is also necessary during G(1) phase (after mitosis). Prolactin can elicit the overt differentiative responses after mitosis. Activity of hydrocortisone precedes that of prolactin, that is, after mitosis it is not capable of eliciting the differentiative response.
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