Interrogating cellular fate decisions with high-throughput arrays of multiplexed cellular communities.

Sisi Chen,Sonny Hsiao,Michel M Maharbiz,Andrew W Bremer,Yun Suk Na,David V Schaffer,Michael E Todhunter,Olivia J Scheideler,Zev J Gartner,Prithvi R Bomdica

doi:10.1038/ncomms10309

Abstract

Recreating heterotypic cell–cell interactions in vitro is key to dissecting the role of cellular communication during a variety of biological processes. This is especially relevant for stem cell niches, where neighbouring cells provide instructive inputs that govern cell fate decisions. To investigate the logic and dynamics of cell–cell signalling networks, we prepared heterotypic cell–cell interaction arrays using DNA-programmed adhesion. Our platform specifies the number and initial position of up to four distinct cell types within each array and offers tunable control over cell-contact time during long-term culture. Here, we use the platform to study the dynamics of single adult neural stem cell fate decisions in response to competing juxtacrine signals. Our results suggest a potential signalling hierarchy between Delta-like 1 and ephrin-B2 ligands, as neural stem cells adopt the Delta-like 1 phenotype of stem cell maintenance on simultaneous presentation of both signals.

Highlights

Recreating heterotypic cell–cell interactions in vitro is key to dissecting the role of cellular communication during a variety of biological processes
Adult neural stem cells (NSCs)[8,9,10] in the brain generate new neurons to modulate learning and memory, a process tightly regulated by a repertoire of neighbouring cells that present a spectrum of signals (Eph-ephrin[11], Notch-Delta[12], Wnt[13], Shh[14] and so on)
Net difference in data the distributions of percent Tuj[1] þ cells per island and the total number of Tuj[1] þ cells produced were similar when NSCs were cultured with an astrocyte expressing Dll[1] alone or a Dll[1] astrocyte plus an EfnB2 astrocyte (Supplementary Fig. 11b). These results suggest that, in dynamic niche microenvironments, competing juxtacrine signals from Dll[1] and EfnB2 may be interpreted by NSCs as a Dll[1] signal

Summary

Introduction

Recreating heterotypic cell–cell interactions in vitro is key to dissecting the role of cellular communication during a variety of biological processes. Networks of interacting cells regulate the biology and pathology of all mammalian tissues, including positive–negative selection in adaptive immune responses[1], tumour–stromal–vascular interactions during cancer progression[2] and stem cell-niche interactions during development and adulthood[3] Within these intercellular signalling networks, the relative number and spatial organization of diverse cell types contributes to the behaviour of the system as a whole[4]. The stem cell niche is an example of a cell community containing a diversity of interacting cells that orchestrate tissue development, maintenance and repair[3] Within this milieu, spatially restricted extracellular signals guide stem cell selfrenewal and differentiation[5]. Microfluidic and micropatterned platforms offer improved throughput and the capacity for single-cell analysis but are typically inefficient because they rely on Poisson statistics to generate arrays of interacting cells, are incapable of robust manipulation of more than two cell types at the single-cell level and restrict cell motility and proliferation[16,17]

Methods

Results

Conclusion