Adhesive forces in embryonic stem cell cultures |
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Authors: | Alicia A Blancas Chi-Shuo Chen Sarah Stolberg Kara E McCloskey |
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Affiliation: | 1.Graduate Program in Quantitative and Systems Biology; University of California Merced; Merced, CA USA;2.Graduate Program in Biological Engineering and Small-Scale Technologies; University of California Merced; Merced, CA USA;3.School of Engineering; University of California Merced; Merced, CA USA |
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Abstract: | Most cell culture systems grow and spread as contact-inhibited monolayers on flat culture dishes, but the embryonic stem cell (ESC) is one of the cell phenotypes that prefer to self-organize as tightly packed three-dimensional (3D) colonies. ESC also readily form 3D cell aggregates, called embryoid bodies (EB) that partially mimic the spatial and temporal processes of the developing embryo. Here, the rationale for ESC aggregation, rather than “spreading” on gelatin-coated or mouse embryonic fibroblast (MEF)-coated dishes, is examined through the quantification of the expression levels of adhesion molecules on ESC and the calculation of the adhesive forces on ESC. Modeling each ESC as a dodecahedron, the adhesive force for each ESC-ESC binding was found to be 9.1 × 105 pN, whereas, the adhesive force for ESC-MEF binding was found to be an order of magnitude smaller at 7.9 × 104 pN. We also show that E-cadherin is the dominating molecule in the ESC-ESC adhesion and blocking E-cadherin leads to a significant reduction in colony formation. Here, we mathematically describe the preference for ESC to self-assemble into ESC-ESC aggregates and 3D colonies, rather than to bind and spread on gelatin or MEF-coated dishes, and have shown that these interactions are predominantly due to E-cadherin expression on ESC.Key words: embryonic stem cells, stem cell morphology, E-cadherin, beta-1 integrin, cell adhesive forces, quantitative flow cytometry |
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