A pH Sensor Scaffold for Mapping Spatiotemporal Gradients in Three Dimensional In Vitro Tumour Models

Riccardo Rizzo; Valentina Onesto; Stefania Forciniti; Anil Chandra; Saumya Prasad; Helena Iuele; Francesco Colella; Giuseppe Gigli; Loretta L. del Mercato

doi:10.1016/j.bios.2022.114401

arXiv · 2403.02838

A pH Sensor Scaffold for Mapping Spatiotemporal Gradients in Three Dimensional In Vitro Tumour Models

Riccardo Rizzo·Valentina Onesto·Stefania Forciniti·Anil Chandra·Saumya Prasad·Helena Iuele·Francesco Colella·Giuseppe Gigli·Loretta L. del Mercato

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Abstract

The detection of extracellular pH at single cell resolution is challenging and requires advanced sensibility. Sensing pH at a high spatial and temporal resolution might provide crucial information in understanding the role of pH and its fluctuations in a wide range of physio-pathological cellular processes, including cancer. Here, a method to embed silica-based fluorescent pH sensors into alginate-based three-dimensional (3D) microgels tumour models, coupled with a computational method for fine data analysis, is presented. By means of confocal laser scanning microscopy, live-cell time-lapse imaging of 3D alginate microgels was performed and the extracellular pH metabolic variations were monitored in both in vitro 3D mono- and 3D co-cultures of tumour and stromal pancreatic cells. The results show that the extracellular pH is cell line-specific and time-dependent. Moreover, differences in pH were also detected between 3D monocultures versus 3D co-cultures, thus suggesting the existence of a metabolic crosstalk between tumour and stromal cells. In conclusion, the system has the potential to image multiple live cells types in a 3D environment and to decipher in real-time their pH metabolic interplay under controlled experimental conditions, thus being also a suitable platform for drug screening and personalized medicine.

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Riccardo Rizzo, Valentina Onesto, Stefania Forciniti, Anil Chandra, Saumya Prasad, Helena Iuele, Francesco Colella, Giuseppe Gigli, Loretta L. del Mercato. 2024-03-05. A pH Sensor Scaffold for Mapping Spatiotemporal Gradients in Three Dimensional In Vitro Tumour Models. https://doi.org/10.1016/j.bios.2022.114401

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