RRC ID 4758
Author Nagaoka M, Hagiwara Y, Takemura K, Murakami Y, Li J, Duncan SA, Akaike T.
Title Design of the artificial acellular feeder layer for the efficient propagation of mouse embryonic stem cells.
Journal J. Biol. Chem.
Abstract Embryonic stem (ES) cells are pluripotent-undifferentiated cells that have a great interest for the investigation of developmental biology. Murine ES cells maintain their pluripotency by the supplementation of the leukemia inhibitory factor (LIF). LIF is reported to act as a matrix-anchored form, and immobilized cytokines are useful to sustain their signaling on target cells. In this study, we used the immobilizable fusion protein composed of LIF and IgG-Fc region, which was used as a model of the matrix-anchored form of LIF to establish a novel system for ES cell culture and to investigate the effect of immobilized LIF on maintenance of ES cell pluripotency. Mouse ES cells maintained their undifferentiated state on the surface coated with LIF-Fc. Furthermore, when cultured on the co-immobilized surface with LIF-Fc and E-cadherin-Fc, mouse ES cells showed characteristic scattering morphologies without colony formation, and they could maintain their undifferentiated state and pluripotency without additional LIF supplementation. The activation of LIF signaling was sustained on the co-immobilized surface. These results indicate that immobilized LIF and E-cadherin can maintain mouse ES cells efficiently and that the immobilizable LIF-Fc fusion protein is useful for the investigation of signaling pathways of an immobilized form of LIF in the maintenance of ES cell pluripotency.
Volume 283(39)
Pages 26468-76
Published 2008-9-26
DOI 10.1074/jbc.M805037200
PII M805037200
PMID 18614540
PMC PMC3258920
MeSH Animals Cadherins / chemistry Cadherins / genetics Cadherins / metabolism Cell Culture Techniques / methods* Cell Line Embryonic Stem Cells / cytology* Extracellular Matrix / chemistry Extracellular Matrix / genetics Extracellular Matrix / metabolism Immunoglobulin Constant Regions / chemistry Immunoglobulin Constant Regions / genetics Immunoglobulin Constant Regions / pharmacology* Leukemia Inhibitory Factor / chemistry Leukemia Inhibitory Factor / genetics Leukemia Inhibitory Factor / pharmacology* Mice Pluripotent Stem Cells / cytology* Recombinant Fusion Proteins / chemistry Recombinant Fusion Proteins / genetics Recombinant Fusion Proteins / pharmacology Signal Transduction / drug effects
IF 4.011
Times Cited 21
WOS Category BIOCHEMISTRY & MOLECULAR BIOLOGY
Resource
DNA material Mouse E-cadherin full length cDNA (RDB01184)