In vitro differentiation of mouse hepatocytes into insulin-producing cells induced by small molecules
LIAO Jing-jing
HAO Hao-jie
LIU Jian-ping
Abstract：Objective To induce differentiation of isolated primary mouse hepatocytes into insulin-producing cells ( IPCs) with small molecules.Methods Mouse hepatocytes were isolated and cultured from minced mice livers be-fore being identified by glycogen staining.They were subsequently treated with the following inducers, 5-aza-2′-de-oxycytidine (5-AZA), richostatin A (TSA), retinoic acid (RA), insulin-transferrin-selenium (ITS), nicotinamide ( NA) and with culture media of different glucose concentration.Hepatocytes cultured without inducers were served as the negative control.Results Primary hepatocytes underwent dedifferentiation in the effect of 5-AZA and TSA.RT-qPCR demonstrated significant decrease of albumin, a surface marker of mature hepatocytes, along with nestin-positive cells. Further culture with low glucose medium containing either RA plus ITS or NA alone for 7 d yielded Pdx1 and insulin posi-tive cells at both gene and protein levels.Conclusion Small molecules (5-AZA, TSA, RA, ITS, NA) and the se-quential use of high and low concentration of glucose can induce hepatocytes into pancreatic precursor cells, and further differentiate into insulin-producing cells in a short period (17 d) .
Keywords：hepatocytessmall moleculesinsulin-producing cellsdiabetes mellitus
Publication Date：2015-07-17
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：5( 2128-2131,2132 )