Spatially resolved analysis of microenvironmental gradient impact on cancer cell phenotypes.
Jamie AuxillosRoxane CrouigneauYan-Fang LiYifan DaiArnaud StiglianiIsabella TavernaroUte Resch-GengerAlbin SandelinRodolphe MarieStine Helene Falsig PedersenPublished in: Science advances (2024)
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.
Keyphrases
- single cell
- high resolution
- rna seq
- high throughput
- cell therapy
- induced apoptosis
- gene expression
- circulating tumor cells
- stem cells
- transcription factor
- squamous cell carcinoma
- genome wide
- papillary thyroid
- endoplasmic reticulum stress
- oxidative stress
- staphylococcus aureus
- cell death
- mesenchymal stem cells
- young adults
- high speed