Isolation and morphologic characterization of bile duct epithelial cells from normal rat liver
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2015, American Journal of PathologyCitation Excerpt :In cholangiocytes from normal rats treated with saline or GnRH we evaluated intracellular cAMP and IP3 levels and the expression of SR, CFTR, and AE2 (functional indices of biliary growth)2,5–7,9 by real-time PCR. After purification, cholangiocytes were incubated for 1 hour at 37°C33 and subsequently incubated for 5 minutes (cAMP) or 10 minutes (IP3) at room temperature before evaluating the levels of these messengers.9,39 To validate our models, we measured GnRH mRNA expression in the hypothalamus and cholangiocytes by real-time PCR, and GnRH levels by EIA kits40 in the serum and the media of primary cultures (after incubation for 6 hours) of cholangiocytes from the selected groups of animals.