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

Int J Dev Biol. 2004 Dec;48(10):1095-104. doi: 10.1387/ijdb.041904pb.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • C-Peptide / chemistry
  • Carbonic Anhydrase II / metabolism
  • Cell Culture Techniques
  • Cell Differentiation
  • Electrophysiology
  • Embryo, Mammalian / cytology*
  • Enzyme-Linked Immunosorbent Assay
  • Homeodomain Proteins / metabolism
  • Insulin / metabolism*
  • Intermediate Filament Proteins / metabolism*
  • Islets of Langerhans / cytology*
  • Islets of Langerhans / embryology*
  • Islets of Langerhans / metabolism
  • Keratins / metabolism
  • Male
  • Mice
  • Mice, Inbred BALB C
  • Microscopy, Fluorescence
  • Models, Biological
  • Nerve Tissue Proteins / metabolism*
  • Nestin
  • Patch-Clamp Techniques
  • Peptides / chemistry
  • Phenotype
  • RNA, Messenger / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Stem Cells / cytology*
  • Streptozocin / pharmacology
  • Time Factors
  • Trans-Activators / metabolism

Substances

  • C-Peptide
  • Homeodomain Proteins
  • Insulin
  • Intermediate Filament Proteins
  • Nerve Tissue Proteins
  • Nes protein, mouse
  • Nestin
  • Peptides
  • RNA, Messenger
  • Trans-Activators
  • pancreatic and duodenal homeobox 1 protein
  • Streptozocin
  • Keratins
  • Carbonic Anhydrase II