title: The self-renewal of mouse embryonic stem cells is regulated by cell-substratum adhesion and cell spreading.
abstract: Mouse embryonic stem cells (mESCs) undergo self-renewal in the presence of the cytokine, leukaemia inhibitory factor (LIF). Following LIF withdrawal, mESCs differentiate, and this is accompanied by an increase in cell-substratum adhesion and cell spreading. The purpose of this study was to investigate the relationship between cell spreading and mESC differentiation. Using E14 and R1 mESC lines, we have restricted cell spreading in the absence of LIF by either culturing mESCs on chemically defined, weakly adhesive biomaterial substrates, or by manipulating the cytoskeleton. We demonstrate that by restricting the degree of spreading by either method, mESCs can be maintained in an undifferentiated and pluripotent state. Under these conditions, self-renewal occurs without the need for LIF and is independent of nuclear translocation of tyrosine-phosphorylated STAT3 or β-catenin, which have previously been implicated in self-renewal. We also demonstrate that the effect of restricted cell spreading on mESC self-renewal is not mediated by increased intercellular adhesion, as evidenced by the observations that inhibition of mESC adhesion using a function blocking anti E-cadherin antibody or siRNA do not promote differentiation. These results show that mESC spreading and differentiation are regulated both by LIF and by cell-substratum adhesion, consistent with the hypothesis that cell spreading is the common intermediate step in the regulation of mESC differentiation by either LIF or cell-substratum adhesion.
authors.affiliation: Institute of Translational Medicine, The University of Liverpool, Liverpool L69 3GE, UK. Electronic address: p.a.murray@liv.ac.uk.
authors.forename: Patricia
authors.initials: P
authors.lastname: Murray
authors.forename: Marina
authors.initials: M
authors.lastname: Prewitz
authors.forename: Isabel
authors.initials: I
authors.lastname: Hopp
authors.forename: Nicola
authors.initials: N
authors.lastname: Wells
authors.forename: Haifei
authors.initials: H
authors.lastname: Zhang
authors.forename: Andrew
authors.initials: A
authors.lastname: Cooper
authors.forename: Kristina L
authors.initials: KL
authors.lastname: Parry
authors.forename: Robert
authors.initials: R
authors.lastname: Short
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Antoine
authors.forename: David
authors.initials: D
authors.lastname: Edgar
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D001672:Biocompatible Materials / Q000494:pharmacology*; D015820:Cadherins / Q000378:metabolism; D002448:Cell Adhesion / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects* / Q000201:enzymology; D022001:Focal Adhesions / Q000187:drug effects / Q000378:metabolism; D053605:Leukemia Inhibitory Factor / Q000494:pharmacology; D051379:Mice; D010766:Phosphorylation / Q000187:drug effects; D050796:STAT3 Transcription Factor / Q000378:metabolism; D022502:Stress Fibers / Q000187:drug effects / Q000378:metabolism; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism
chemical_list: D000199:Actins; D001672:Biocompatible Materials; D015820:Cadherins; D053605:Leukemia Inhibitory Factor; D050796:STAT3 Transcription Factor; D054460:rho-Associated Kinases
keywords: Biomaterials; Cell spreading; E-cadherin; Embryonic stem cells; FAK; LIF; PLGA; ROCK; Rho kinase; focal adhesion kinase; leukaemia inhibitory factor; mESC; mouse embryonic stem cell; pPAA; plasma polymerised acrylic acid; poly(lactic-co-glycolic acid); rho kinase