期刊
BIOSENSORS & BIOELECTRONICS
卷 212, 期 -, 页码 -出版社
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2022.114401
关键词
Optical pH-sensors; Alginate microgels; In vitro 3D tumour models; Tumour microenvironment; Cell metabolism; Computational analysis
类别
资金
- European Research Council (ERC) under the European Union's Horizon 2020 research and innovation program ERC Starting Grant INTERCELLMED [759959]
- My First AIRC Grant [22902]
- PRIN-2022 [20205B2HZE_004]
- Tecnopolo per la medicina di precisione (TecnoMed Puglia) Regione Puglia: DGR [2117, CUP: B84I18000540002]
This study presents a method to embed fluorescent pH sensors into 3D tumor models and analyze the data using computational methods. The results show that extracellular pH is cell line-specific and time-dependent. Additionally, metabolic crosstalk between tumor cells and stromal cells was detected.
The detection of extracellular pH at single cell resolution is challenging and requires advanced sensibility. Sensing pH at 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 cell 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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