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These 5 factors permit growth for one month in the absence of serum, and a growth rate equivalent to that observed in serum\u2010supplemented medium. Dibutyryl cAMP substitutes for prostaglandin E<jats:sub>1<\/jats:sub> in the medium, suggesting that increased growth of Maden Darby canine kidney cells results from increased intracellular cAMP. Potential applications of the serum\u2010free medium are discussed. 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