Neural crest cell induction and cell shape changes on biofunctionalized lipid bilayers

Neural crest cells (NCC) are a multipotent cell population that displays a high migratory potential. For initiation of motility structures and pathfinding during the migration NCC form contacts via cell-cell adhesion molecules like cadherin-11. To investigate the morphogenetic response of NCC to such a cell-cell contact we make use of artificial two-dimensional lipid membranes biofunctionalized with tailor-made recombinant Xenopus Cadherin-11. The aim is to give a cue what guides morphogenetic cell changes in NCC-induced animal cap tissue in a quantitative manner.

In the first step we established the parameters necessary for neural crest induction in naïve animal cap tissue and long-term cultivation on functionalized lipid bilayers. We injected Xenopus embryos at the 2-cell-stage with BMP inhibitor and Wnt activator. At early gastrulation we explanted animal caps of the injected embryos. In case of successful NCC induction these cells transformed into neural crest tissue what was shown by expression of neural crest specific markers, changes of explant morphology, and changes in cell polarity from epithelial to front-tail migrating manner shown by EB1-eGFP movement.

We showed the formation and biofunctionalization of a lipid bilayer via QCM-D measurements. A linear increase of adsorbed mass as a function of anchor (DOGS-NTA) density was observed. We also used RICM to demonstrate the adhesion of animal caps explants to the Xcad-11 functionalized bilayers.