Abstract
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.
Highlights
The genes and surrounding environment of a cell regulate its function
We explore the application of our laser-guided cell micropatterning (LGCM) system[15] in combination with surface patterning methods[16] to investigate stem cell differentiation at the singlecell level in a cardiomyocyte microculturing environment
When the beam waist was as large as 4 mm, the axial optical force remains positive along the axial direction in all longitudinal sections (Figure 3c), forming the pure-guidance mode
Summary
The genes and surrounding environment of a cell regulate its function. The interplay between the complex process of gene expression and microenvironmental intricacies causes large functional variation, even among cells within a small region of tissue. Bio-MEMS techniques have been developed to create an engineered cell culture for systematic observation of the interactions of cells with their surroundings, including interactions with the adjacent extracellular matrix.[5] laser single-cell microdissection techniques have been developed for harvesting single cells from the culturing environment for various biological assays.[6] Further, various single-cell polymerase chain reaction (PCR) techniques[7] are capable of assessing gene expression intercellular variability and correlating the result to the specific microenvironments.[8]
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