Embryonic stem cells differentiate into insulin-producing cells without selection of nestin-expressing cells.
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.
Blyszczuk
Przemyslaw
P
In Vitro Differentiation Group, Institute of Plant Genetics and Crop Plant Research (IPK), Gatersleben, Germany.
Asbrand
Christian
C
Rozzo
Aldo
A
Kania
Gabriela
G
St-Onge
Luc
L
Rupnik
Marjan
M
Wobus
Anna M
AM
D000818:Animals; D002096:C-Peptide / Q000737:chemistry; D024402:Carbonic Anhydrase II / Q000378:metabolism; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D004594:Electrophysiology; D004622:Embryo, Mammalian / Q000166:cytology*; D004797:Enzyme-Linked Immunosorbent Assay; D018398:Homeodomain Proteins / Q000378:metabolism; D007328:Insulin / Q000378:metabolism*; D007381:Intermediate Filament Proteins / Q000378:metabolism*; D007515:Islets of Langerhans / Q000166:cytology* / Q000196:embryology* / Q000378:metabolism; D007633:Keratins / Q000378:metabolism; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008856:Microscopy, Fluorescence; D008954:Models, Biological; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D064231:Nestin; D018408:Patch-Clamp Techniques; D010455:Peptides / Q000737:chemistry; D010641:Phenotype; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013234:Stem Cells / Q000166:cytology*; D013311:Streptozocin / Q000494:pharmacology; D013997:Time Factors; D015534:Trans-Activators / Q000378:metabolism
D002096:C-Peptide; D018398:Homeodomain Proteins; D007328:Insulin; D007381:Intermediate Filament Proteins; D009419:Nerve Tissue Proteins; C577485:Nes protein, mouse; D064231:Nestin; D010455:Peptides; D012333:RNA, Messenger; D015534:Trans-Activators; C084010:pancreatic and duodenal homeobox 1 protein; D013311:Streptozocin; D007633:Keratins; D024402:Carbonic Anhydrase II