Large-scale patterning of single cells and cell clusters in hydrogels

Abstract

Biophysical properties of the extracellular matrix (ECM) are known to play a significant role in cell behavior. To gain a better understanding of the effects of the biophysical microenvironment on cell behavior, the practical challenge is longitudinally monitoring behavioral variations within a population to make statistically powerful assessments. Population-level measurements mask heterogeneity in cell responses, and large-scale individual cell measurements are often performed in a one-time, snapshot manner after removing cells from their matrix. Here we present an easy and low-cost method for large-scale, longitudinal studies of heterogeneous cell behavior in 3D hydrogel matrices. Using a platform we term “the drop-patterning chip”, thousands of cells were simultaneously transferred from microwell arrays and fully embedded, only using the force of gravity, in precise patterns in 3D collagen I or Matrigel. This method allows for throughputs approaching 2D patterning methods that lack phenotypic information on cell-matrix interactions, and does not rely on special equipment and cell treatments that may result in a proximal stiff surface. With a large and yet well-organized group of cells captured in 3D matrices, we demonstrated the capability of locating selected individual cells and monitoring cell division, migration, and proliferation for multiple days.

Multiplexing physical stimulation on single human induced pluripotent stem cell-derived cardiomyocytes for phenotype modulation

“…Engineering surface topography to provide a niche that mimics the native context of cells is being actively exploited for biomedical research, including tissue engineering using single and/or multiple cell types [59]. Previously, entrapment of single cells and/or cell clusters with 3D micropatterns was piloted to be used in cancer research [60] and drug screening [61] with very limited use in the cardiac field. Previous studies showed the relationship between biophysical cues and CMs in many settings.…”

High‐Precision 3D Bio‐Dot Printing to Improve Paracrine Interaction between Multiple Types of Cell Spheroids

“…[ 9 ] However, this technique allows the cell spheroids to be arranged exclusively in a 2D structure, and often with only a single type of spheroid. [ 7,10 ] Alternatively, a magnetic field‐based technique has been introduced to create patterns with cell aggregates. [ 11 ] Although this technique allows the patterned spheroids to be induced within a short time in a 3D environment, several researchers have reported that the integrity of the cell aggregates is reduced when the field disappears; moreover, the technique cannot produce patterns that incorporate multiple cell aggregates.…”

Mechanophenotyping of 3D multicellular clusters using displacement arrays of rendered tractions

“…In particular, the epithelial-mesenchymal transition (EMT) is associated with weakened cell-cell junctions and increased cellmatrix adhesions, driving tissue disorganization and dissemination (2). Remarkably, multicellular clusters cultured ex vivo in compliant 3D hydrogels can exhibit tissue-like form and function (3)(4)(5)(6)(7), representing a promising preclinical testbed for higher-throughput drug discovery and development (8,9). However, existing assays for 3D mechanophenotyping have been limited to a few cells per experiment due to the need for high-resolution optics and labor-intensive image processing, as well as complex readouts of force generation (10).…”