Laser patterning for the study of MSC cardiogenic differentiation at the single-cell level

Mesenchymal stem cells (MSCs) have been cited as contributors to heart repair through cardiogenic differentiation and multiple cellular interactions, including the paracrine effect, cell fusion, and mechanical and electrical couplings. Due to heart–muscle complexity, progress in the development of knowledge concerning the role of MSCs in cardiac repair is heavily based on MSC–cardiomyocyte coculture. In conventional coculture systems, however, the in vivo cardiac muscle structure, in which rod-shaped cells are connected end-to-end, is not sustained; instead, irregularly shaped cells spread randomly, resulting in randomly distributed cell junctions. Consequently, contact-mediated cell–cell interactions (e.g., the electrical triggering signal and the mechanical contraction wave that propagate through MSC–cardiomyocyte junctions) occur randomly. Thus, the data generated on the beneficial effects of MSCs may be irrelevant to in vivo biological processes. In this study, we explored whether cardiomyocyte alignment, the most important phenotype, is relevant to stem cell cardiogenic differentiation. Here, we report (i) the construction of a laser-patterned, biochip-based, stem cell–cardiomyocyte coculture model with controlled cell alignment; and (ii) single-cell-level data on stem cell cardiogenic differentiation under in vivo-like cardiomyocyte alignment conditions. Bruce Gao from Clemson University and collaborators from the USA and China used optical forces from a laser beam to deposit a single stem cell in a microengineered cardiomyocyte culture that mimics in vivo cell alignment. The cell was then harvested from its niche and analyzed at the single-cell level for its differentiation status. Although other methods have achieved similar behaviour for larger groups of cells, this laser-based technique allows the study of single stem-cell differentiation, which is critical for understanding the fundamental aspects of heart cell development.