Dimeric integrin a5ß1 ligands confer morphological and differentiation responses to murine embryonic stem cells

TY - JOUR

T1 - Dimeric integrin a5ß1 ligands confer morphological and differentiation responses to murine embryonic stem cells

AU - Singh, M.D.

AU - Kreiner, M.

AU - McKimmie, C.S.

AU - Holt, S.

AU - van der Walle, C.F.

AU - Graham, G.J.

PY - 2009/12/18

Y1 - 2009/12/18

N2 - We present the first report utilizing, and showing the functional relevance of, self-assembling polyvalent ligands specific for integrin α5β1 in murine embryonic stem (mES) cell adhesion. Di, tri and tetrameric 9th-10th type III fibronectin domains (FIII9′10) were used to generate clustered integrin α5β1 ligand surfaces for mES cell culture. Compared to gelatin, FIII9′10 (monomer), FIII9′10-trimer and -tetramer, the FIII9′10-dimer supported the highest number of mES cell colonies. No evidence of domain unfolding upon surface adsorption was found. Colonies appeared disperse with a spread cell morphology unless subdued back to a tight morphology with increasing concentrations of leukemia inhibitory factor (LIF). In the presence of LIF, mES cells adherent to the FIII9′10-dimer showed transient upregulation of Oct-4, the mesodermal transcription factor, Brachyury, and the ectodermal marker, Nestin. However, dual upregulation of Nanog maintained the mES cells in a pluripotent state, confirmed by alkaline phosphatase staining. Therefore, the behavior of mES cells adherent to dimeric integrin α5β1 ligands is a largely morphological phenomenon conferring pro-differentiation signals towards mesodermal and ectodermal lineages. This work will be of interest to cell and tissue engineering groups aiming to control ES cell behavior through integrin ligand presentation and synthetic substrates.

AB - We present the first report utilizing, and showing the functional relevance of, self-assembling polyvalent ligands specific for integrin α5β1 in murine embryonic stem (mES) cell adhesion. Di, tri and tetrameric 9th-10th type III fibronectin domains (FIII9′10) were used to generate clustered integrin α5β1 ligand surfaces for mES cell culture. Compared to gelatin, FIII9′10 (monomer), FIII9′10-trimer and -tetramer, the FIII9′10-dimer supported the highest number of mES cell colonies. No evidence of domain unfolding upon surface adsorption was found. Colonies appeared disperse with a spread cell morphology unless subdued back to a tight morphology with increasing concentrations of leukemia inhibitory factor (LIF). In the presence of LIF, mES cells adherent to the FIII9′10-dimer showed transient upregulation of Oct-4, the mesodermal transcription factor, Brachyury, and the ectodermal marker, Nestin. However, dual upregulation of Nanog maintained the mES cells in a pluripotent state, confirmed by alkaline phosphatase staining. Therefore, the behavior of mES cells adherent to dimeric integrin α5β1 ligands is a largely morphological phenomenon conferring pro-differentiation signals towards mesodermal and ectodermal lineages. This work will be of interest to cell and tissue engineering groups aiming to control ES cell behavior through integrin ligand presentation and synthetic substrates.

KW - embryonic stem cells

KW - cell adhesion

KW - fibronectin

KW - integrin

KW - self-assembly

KW - neutron reflectivity

UR - http://dx.doi.org/10.1016/j.bbrc.2009.10.035

U2 - 10.1016/j.bbrc.2009.10.035

DO - 10.1016/j.bbrc.2009.10.035

M3 - Article

VL - 390

SP - 716

EP - 721

JO - Biochemical and Biophysical Research Communications

T2 - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

SN - 1090-2104

IS - 3

ER -