TY - JOUR
T1 - Fibronectin-mediated upregulation of α5β1 integrin and cell adhesion during differentiation of mouse embryonic stem cells
AU - Pimton, Pimchanok
AU - Sarkar, Saheli
AU - Sheth, Nidhi
AU - Perets, Anat
AU - Marcinkiewicz, Cezary
AU - Lazarovici, Philip
AU - Lelkes, Peter I.
PY - 2011
Y1 - 2011
N2 - Embryonic stem (ES) cells have a broad potential application in regenerative medicine and can be differentiated into cells of all three germ layers. Adhesion of es cells to extracellular matrix (eCM) proteins is essential for the differentiation pathway; Cell-eCM adhesion is mediated by integrins that have the ability to activate many intracellular signaling pathways. Therefore, we hypothesize that the expression and function of integrin receptors is a critical step in es differentiation. Using functional cell adhesion assays, our study demonstrates that α5β1 is a major functional integrin receptor expressed on the cell surface of undifferentiated mouse es-D3 cells, which showed significantly higher binding to fibronectin as compared to collagens. This adhesion was specific mediated by integrin α5β1 as evident from the inhibition with a disintegrin selective for this particular integrin. Differentiation of es-D3 cells on fibronectin or on a collagen type1/fibronectin matrix, caused further selective upregulation of the α5β1 integrin. Differentiation of the cells, as evaluated by immunofluorescence, FACs analysis and quantitative RT-pCR, was accompanied by the upregulation of mesenchymal (Flk1, isolectin B4, α-sMA, vimentin) and endodermal markers (FoxA2, sOX 17, cytokeratin) in parallel to increased expression of α5β1 integrin. Taken together, the data indicate that fibronectin-mediated, upregulation of α5β1 integrin and adhesion of es-D3 cells to specific eCM molecules are linked to early stages of mouse embryonic stem cells commitment to meso-endodermal differentiation.
AB - Embryonic stem (ES) cells have a broad potential application in regenerative medicine and can be differentiated into cells of all three germ layers. Adhesion of es cells to extracellular matrix (eCM) proteins is essential for the differentiation pathway; Cell-eCM adhesion is mediated by integrins that have the ability to activate many intracellular signaling pathways. Therefore, we hypothesize that the expression and function of integrin receptors is a critical step in es differentiation. Using functional cell adhesion assays, our study demonstrates that α5β1 is a major functional integrin receptor expressed on the cell surface of undifferentiated mouse es-D3 cells, which showed significantly higher binding to fibronectin as compared to collagens. This adhesion was specific mediated by integrin α5β1 as evident from the inhibition with a disintegrin selective for this particular integrin. Differentiation of es-D3 cells on fibronectin or on a collagen type1/fibronectin matrix, caused further selective upregulation of the α5β1 integrin. Differentiation of the cells, as evaluated by immunofluorescence, FACs analysis and quantitative RT-pCR, was accompanied by the upregulation of mesenchymal (Flk1, isolectin B4, α-sMA, vimentin) and endodermal markers (FoxA2, sOX 17, cytokeratin) in parallel to increased expression of α5β1 integrin. Taken together, the data indicate that fibronectin-mediated, upregulation of α5β1 integrin and adhesion of es-D3 cells to specific eCM molecules are linked to early stages of mouse embryonic stem cells commitment to meso-endodermal differentiation.
KW - Adhesion
KW - Differentiation
KW - Disintegrins
KW - Fibronectin
KW - LIF
KW - Mouse embryonic stem cells
KW - Upregulation
KW - α5β1 integrin
UR - http://www.scopus.com/inward/record.url?scp=78650832005&partnerID=8YFLogxK
U2 - 10.4161/cam.5.1.13704
DO - 10.4161/cam.5.1.13704
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AN - SCOPUS:78650832005
SN - 1933-6918
VL - 5
SP - 73
EP - 82
JO - Cell Adhesion and Migration
JF - Cell Adhesion and Migration
IS - 1
ER -