Micropatterned mammalian cells exhibit phenotype-specific left-right asymmetry

Abstract

Left-right (LR) asymmetry (handedness, chirality) is a well-conserved biological property of critical importance to normal development. Changes in orientation of the LR axis due to genetic or environmental factors can lead to malformations and disease. While the LR asymmetry of organs and whole organisms has been extensively studied, little is known about the LR asymmetry at cellular and multicellular levels. Here we show that the cultivation of cell populations on micropatterns with defined boundaries reveals intrinsic cell chirality that can be readily determined by image analysis of cell alignment and directional motion. By patterning 11 different types of cells on ring-shaped micropatterns of various sizes, we found that each cell type exhibited definite LR asymmetry (p value down to 10 −185 ) that was different between normal and cancer cells of the same type, and not dependent on surface chemistry, protein coating, or the orientation of the gravitational field. Interestingly, drugs interfering with actin but not microtubule function reversed the LR asymmetry in some cell types. Our results show that micropatterned cell populations exhibit phenotype-specific LR asymmetry that is dependent on the functionality of the actin cytoskeleton. We propose that micropatterning could potentially be used as an effective in vitro tool to study the initiation of LR asymmetry in cell populations, to diagnose disease, and to study factors involved with birth defects in laterality.cell patterning | tissue morphogenesis | cell polarity | cell migration

Emergence of collective modes and tri-dimensional structures from epithelial confinement

“…Of note, clockwise rotation was slightly but significantly favoured compared with counter-clockwise ( Supplementary Fig. 6), as already observed with other cell types 23 . This asymmetry is likely to result from intrinsic cell chirality; it is revealed here because of the relatively small and thus coherently moving cell population in presence of the adhesive/non-adhesive boundary.…”

Cell Chirality Induces Collective Cell Migration in Epithelial Sheets

“…Chiral behavior of cells has been reported elsewhere recently [25][26][27] and has been linked to chirally asymmetric dynamics of parts of the cytoskeleton, such as the actomyosin cortex beneath cell-surface membranes [28,29]. More recently, chiral mechanical torque generation in cells has been reported in C. elegans, which contributes to the formation of the left-right body axis [30].…”

Configurable 2D and 3D spheroid tissue cultures on bioengineered surfaces with acquisition of epithelial–mesenchymal transition characteristics

“…For example, some techniques typically require a substantial amount of cells to seed onto open surfaces and wash away numerous unattached cells after the cell-patterning step has been executed. 1,8,14,18 These result in cell waste and restrict potential applications, for example, rare cells 19 and patient samples. 10 In addition, complexity arises from the use of external systems, such as syringe pumps or electric power, which is less attractive than more simplistic approaches.…”

“…These surfaces have served as versatile tools to explore fundamental cell biology, tissue engineering and drug development, and thereby became key components in a wide range of applications. 10 Various techniques for patterning cells in 2D or 3D tissue culture have demonstrated the ability to direct cells onto selected areas of a surface, including microcontact printing 8 or soft lithography, 1 microfluidic-or microstructure-based methods, 11,12 active control of cell attachment or suspension by physical forces [13][14][15] and layer-bylayer assembly. 16,17 However, challenges still remain.…”

“…[1][2][3][4][5][6] Microsystems open doors to new high-throughput and highcontent approaches, and provide systematic insights into cellular functions influenced by microenvironmental stimuli, which include the interactions between neighboring cells, extracellular matrices, chemical factors and physical forces. [7][8][9] To create and recapitulate the cellular in vivo conditions, biomaterial-based surfaces capable of generating precise patterning of cells for two-dimensional (2D) or three-dimensional (3D) cell culture, and for controlling microenvironmental stimuli at the single-cell level (approximately 10 mm) in microsystems are crucial. These surfaces have served as versatile tools to explore fundamental cell biology, tissue engineering and drug development, and thereby became key components in a wide range of applications.…”