TY - JOUR

T1 - Spatially resolved analysis of microenvironmental gradient impact on cancer cell phenotypes

AU - Auxillos, Jamie

AU - Crouigneau, Roxane

AU - Li, Yan Fang

AU - Dai, Yifan

AU - Stigliani, Arnaud

AU - Tavernaro, Isabella

AU - Resch-Genger, Ute

AU - Sandelin, Albin

AU - Marie, Rodolphe

AU - Pedersen, Stine F.

PY - 2024

Y1 - 2024

N2 - Despite the physiological and pathophysiological significance of microenvironmental gradients, e.g., for diseases such as cancer, tools for generating such gradients and analyzing their impact are lacking. Here, we present an integrated microfluidic-based workflow that mimics extracellular pH gradients characteristic of solid tumors while enabling high-resolution live imaging of, e.g., cell motility and chemotaxis, and preserving the capacity to capture the spatial transcriptome. Our microfluidic device generates a pH gradient that can be rapidly controlled to mimic spatiotemporal microenvironmental changes over cancer cells embedded in a 3D matrix. The device can be reopened allowing immunofluorescence analysis of selected phenotypes, as well as the transfer of cells and matrix to a Visium slide for spatially resolved analysis of transcriptional changes across the pH gradient. This workflow is easily adaptable to other gradients and multiple cell types and can therefore prove invaluable for integrated analysis of roles of microenvironmental gradients in biology.

AB - Despite the physiological and pathophysiological significance of microenvironmental gradients, e.g., for diseases such as cancer, tools for generating such gradients and analyzing their impact are lacking. Here, we present an integrated microfluidic-based workflow that mimics extracellular pH gradients characteristic of solid tumors while enabling high-resolution live imaging of, e.g., cell motility and chemotaxis, and preserving the capacity to capture the spatial transcriptome. Our microfluidic device generates a pH gradient that can be rapidly controlled to mimic spatiotemporal microenvironmental changes over cancer cells embedded in a 3D matrix. The device can be reopened allowing immunofluorescence analysis of selected phenotypes, as well as the transfer of cells and matrix to a Visium slide for spatially resolved analysis of transcriptional changes across the pH gradient. This workflow is easily adaptable to other gradients and multiple cell types and can therefore prove invaluable for integrated analysis of roles of microenvironmental gradients in biology.

U2 - 10.1126/sciadv.adn3448

DO - 10.1126/sciadv.adn3448

M3 - Journal article

C2 - 38701211

AN - SCOPUS:85192133667

VL - 10

JO - Science advances

JF - Science advances

SN - 2375-2548

IS - 18

M1 - eadn3448

ER -