Undersulfation of heparan sulfate restricts differentiation potential of mouse embryonic stem cells.
Heparan sulfate proteoglycans, present on cell surfaces and in the extracellular matrix, interact with growth factors and morphogens to influence growth and differentiation of cells. The sulfation pattern of the heparan sulfate chains formed during biosynthesis in the Golgi compartment will determine the interaction potential of the proteoglycan. The glucosaminyl N-deacetylase/N-sulfotransferase (NDST) enzymes have a key role during biosynthesis, greatly influencing total sulfation of the heparan sulfate chains. The differentiation potential of mouse embryonic stem cells lacking both NDST1 and NDST2 was studied using in vitro differentiation protocols, expression of differentiation markers, and assessment of the ability of the cells to respond to growth factors. The results show that NDST1 and NDST2 are dispensable for mesodermal differentiation into osteoblasts but necessary for induction of adipocytes and neural cells. Gene expression analysis suggested a differentiation block at the primitive ectoderm stage. Also, GATA4, a primitive endoderm marker, was expressed by these cells. The addition of FGF4 or FGF2 together with heparin rescued the differentiation potential to neural progenitors and further to mature neurons and glia. Our results suggest that the embryonic stem cells lacking both NDST1 and NDST2, expressing a very low sulfated heparan sulfate, can take the initial step toward differentiation into all three germ layers. Except for their potential for mesodermal differentiation into osteoblasts, the cells are then arrested in a primitive ectoderm and/or endoderm stage.
Department of Medical Biochemistry and Microbiology, Science for Life Laboratory, Uppsala University, Box 582, 751 23 Uppsala, Sweden.
Maud
M
Forsberg
Katarina
K
Holmborn
Soumi
S
Kundu
Anders
A
Dagälv
Lena
L
Kjellén
Karin
K
Forsberg-Nilsson
D017667:Adipocytes / Q000166:cytology; D000581:Amidohydrolases / Q000172:deficiency* / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D019485:Bone Morphogenetic Proteins / Q000378:metabolism; D002352:Carrier Proteins / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D004475:Ectoderm / Q000166:cytology; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D005346:Fibroblast Growth Factors / Q000494:pharmacology; D055786:Gene Knockout Techniques; D006493:Heparin / Q000494:pharmacology; D006497:Heparan Sulfate / Q000378:metabolism*; D008648:Mesoderm / Q000166:cytology; D051379:Mice; D009154:Mutation; D010006:Osteoblasts / Q000166:cytology; D015398:Signal Transduction / Q000187:drug effects; D015238:Sulfotransferases / Q000172:deficiency* / Q000235:genetics / Q000378:metabolism*; D000099176:Noggin Protein
D000581:Amidohydrolases; D019485:Bone Morphogenetic Proteins; D002352:Carrier Proteins; D005346:Fibroblast Growth Factors; D006493:Heparin; D006497:Heparan Sulfate; D015238:Sulfotransferases; D000099176:Noggin Protein; C491855:Ndst2 protein, mouse; C021350:heparitin sulfotransferase