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Single-Cell Biophysics: Measurement, Modulation, and Modeling
Poster Abstracts
83
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.