Author/Editor		Blyszczuk, Przemyslaw; Asbrand, Christian; Rozzo, Aldo; Kania, Gabriela; St-Onge, Luc; Rupnik, Marjan; Wobus, Anna M
Title		Embryonic stem cells differentiate into-insulin-producting cells without selection of nestin-expressing cells
Type		članek
Source		Int J Dev Biol
Vol. and No.		Letnik 48
Publication year		2004
Volume		str. 1095-104
Language		eng
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 coexpressed 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.
Descriptors		PANCREAS INSULIN C-PEPTIDE STEM CELLS CELL DIFFERENTIATION TRANSCRIPTION, GENETIC POLYMERASE CHAIN REACTION ENZYME-LINKED IMMUNOSORBENT ASSAY PATCH-CLAMP TECHNIQUES FLUORESCENT ANTIBODY TECHNIQUE MICE, INBRED NOD