Single-Cell Biophysics: Measurement, Modulation, and Modeling

Single-Cell Biophysics: Measurement, Modulation, and Modeling

Poster Abstracts

77-POS Board 39 A Scalable, DNA-Based Multicomponent Patterning Method to Model Multivariable Neural Stem Cell-Niche Interactions from a Single-Cell Perspective

Olivia Scheideler , Chun Yang, David Schaffer, Lydia Sohn. University of California, Berkeley, Berkeley, CA, USA.

Biological processes are regulated by complex signaling networks that are challenging to dissect due to the multitude of extrinsic signals that coordinate to guide cell behavior. Understanding these extensive regulatory networks requires not only identifying contributing signaling components – including ligands that are soluble, presented from the extracellular matrix, or neighboring cell surfaces – but also investigating potential synergies or hierarchies between multiple components. In order to enable studies of the later, we have developed a broadly applicable high-throughput, high-resolution DNA-based patterning method that we employ to recapitulate multivariable cell-ligand signaling scenarios. To complement bulk approaches that offer lower resolution, population-level estimates of cell response, our platform offers the unique dual capability to recapitulate cell-cell interactions with single-cell resolution and enable precise spatial control of biologically-relevant “solid-phase” matrix cues. Using photolithographic techniques, we generate multicomponent DNA patterns with spatial and hierarchical complexity across different length scales. We demonstrate that these DNA patterns can instruct the organization of heterogeneous cell populations as well as immobilize multiple ligands with controlled spatial presentations. To demonstrate our method’s unique ability to address complex biological questions, we reconstructed in vitro multifaceted signaling scenarios present within the adult neural stem cell (NSC) niche. Specifically, we generated large-scale arrays of three- component DNA patterns, where one DNA strand encodes the capture of single NSCs and the other two dictates the spatial presentation of two immobilized niche ligands, fibroblast growth factor and an ephrin-B2-mimetic peptide, that are known to promote opposing cell fates. We demonstrate the ability to vary independently the concentration and spatial organization of these two ligands, study their combined effects on single NSC differentiation after long-term culture, and map out the hierarchy of these two signals within the niche.

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