Int. J. Dev. Biol. 48: 1095 - 1104 (2004)
doi: 10.1387/ijdb.041904pb
© UBC Press

Embryonic stem cells differentiate into insulin-producing cells without selection of nestin-expressing cells

Przemyslaw Blyszczuk1, Christian Asbrand2, Aldo Rozzo3, Gabriela Kania1, Luc St-Onge2, Marjan Rupnik3 and Anna M. Wobus*,1

1In Vitro Differentiation Group, Institute of Plant Genetics and Crop Plant Research (IPK), Gatersleben, 2DeveloGen AG, Gttingen and 3European Neuroscience Institute, Gttingen, Germany

ABSTRACT We present a new strategy for the differentiation of embryonic stem (ES) cells into insulin-producing cells via a multi-step process without selection and induction of nestin-positive cells. During ES cell differentiation, transcript levels of genes characteristic of early and mature beta cells including Pdx1, Pax4, insulin and islet amyloid pancreatic peptide are up regulated. Islet-like clusters are characterized by expression of C-peptide, insulin and partially cytokeratin 19 as well as by ion channel activity similar to that found in embryonic beta cells. Cells of islet-like clusters show glucose-dependent insulin release at terminal stage. At an intermediate stage, nestin is partially co-expressed with C-peptide and cytokeratin 19, whereas islet-like clusters at the terminal stage are nestin-negative. We conclude that expression of nestin and cytokeratin 19 is a normal property of ES cells preceding differentiation into C-peptide/insulin-producing cells without any selection for nestin-positive phenotypes.

Keywords:

mouse embryonic stem cell, differentiation, C-peptide, insulin-producing cell, nestin

*Corresponding author e-mail: wobusam@ipk-gatersleben.de