Mir 125b 5p inhibitor

Freshly isolated primary human hepatocytes were provided by Regenerative Medicine and Experimental Surgery, Hannover Medical School, Hannover, as reported.⁽¹²⁾ Mouse primary hepatocytes were isolated from mouse liver as described.⁽⁸⁾ Briefly, mice were anesthetized, and their livers were perfused with Liberase (Roche). After perfusion, livers were disintegrated mechanically before collecting hepatocytes by low‐speed centrifugation. Nonparenchymal cells were removed by discarding the supernatant. For all in vitro transfection experiments, we used Percoll density gradient‐purified mouse hepatocytes to achieve high transfection efficiency. We seeded 100,000 primary hepatocytes per well of a collagen‐coated 12‐well plate (TPP). Twelve hours after seeding, hepatocytes were transfected with 25 nM or 50 nM miR‐125b‐5p mimic, miR‐125b‐5p inhibitor, control scramble (Qiagen), or 100 nM ABTB1 small interfering RNA (siRNA) (Qiagen) using Targefect reagent in the presence of virofect enhancer (Targeting Systems). Transfected hepatocytes were cultured in hepatocyte culture medium (Lonza) containing recombinant epidermal growth factor (an inducer of hepatocyte proliferation), transferrin, ascorbic acid, insulin, hydrocortisone, bovine serum albumin, and gentamicin sulfate‐amphotericin.