NEVADA · Novel microengineered environments for mouse embryonic stem cell (mESC) differentiation towards cardiomyocytes

Novel microengineered environments for mouse embryonic stem cell (mESC) differentiation towards cardiomyocytes

Many forms of heart disease are associated with a decrease in the number of functional cardiomyocytes (CMs). Since the proliferative capacity of CMs decreases significantly in postnatal life, elucidation of the molecular mechanisms that govern cell pluripotent state and lineage commitment in CM renewal bring an additional avenue for therapeutic interventions and disease modeling. Mouse embryonic stem cells (mESCs) constitute an excellent model system for studying these mechanisms due to the availability of various protocols for differentiation and the relative simplicity for genetic manipulation. However, heterogeneity during mESC differentiation has posed a particular problem in the formation of lineage-specific cell populations. Most of the currently employed approaches in cell- and tissue-based engineering studies still involve two-dimensional (2D) surfaces, or monolayer cell cultures, that offer unnatural growth kinetics and cell attachments. However, the monolayer nature of these cultures does not permit cells to grow and proliferate in realistic three-dimensional (3D) microenvironments. 3D synthetic microscaffolds, fabricated by two-photon polymerization (2-PP) photolithography, offer favorable cell responses due to tunable chemical, physical and mechanical properties. 2-PP technology allows the fabrication of volumetric structures of arbitrary shape by directly writing the intended geometry within a photosensitive material. Due to the unprecedented flexibility of 2-PP, matrix architecture and pore size can be controlled with a nanometer resolution. Indeed, none of the currently available microfabrication approaches has been able to investigate cell mechanical properties in 3D with the accuracy of 2-PP photolithography. Chemically defined media combined with the 3D architectures, which more accurately resemble the extracellular environment, offer a powerful tool to mimic specific tissues in vitro. The scientific objective for the project was to establish an advanced cardiomyocytes model to study mESC cardiac differentiation in a biomimetic three-dimensional (3D) environment. Two goals were proposed for this project: • Design of a 3D tailored conductive microscaffold to facilitate spatial cell spreading and to achieve precise stimulation patterns to individual cells in vitro. • Precise quantification of the molecular mechanisms underlying mESC pluripotent state and cardiac differentiation under mechanical and/or electrical stimulation.

Data: CORDIS, © European Union