Real-time and label-free impedimetric analysis of the formation and drug testing of tumor spheroids formed via the liquid overlay technique

To achieve high clinical relevance of drug testing, a spheroid culture model is generally used for a standard assay of early tumors. Currently, the quantification of tumor spheroids is generally based on imaging techniques and conventional bioassays. However, additional processing and sophisticated instrumentation are required, which makes high throughput screening infeasible. In this study, tumor spheroids were formed on a thin layer of non-adherent surface (agarose hydrogel), and their responses were monitored by impedance measurements across the interdigitated electrodes fabricated below the hydrogel. The impedance magnitude change was defined as an index to describe the summation effect of the number and size of tumor spheroids. Real-time and label-free impedimetric analysis of the formation and chemosensitivity of tumor spheroids was demonstrated. The technique has the simplicity of device fabrication and experimental processing. Real-time and quantitative assessment could be realized for a routine tool of a spheroid culture.

[1]  Chia-Hao Huang,et al.  Quantitative impedimetric monitoring of cell migration under the stimulation of cytokine or anti-cancer drug in a microfluidic chip. , 2015, Biomicrofluidics.

[2]  C. Verbeke,et al.  3D pancreatic carcinoma spheroids induce a matrix-rich, chemoresistant phenotype offering a better model for drug testing , 2013, BMC Cancer.

[3]  P. Darbre,et al.  Effect of aluminium on migratory and invasive properties of MCF-7 human breast cancer cells in culture. , 2013, Journal of inorganic biochemistry.

[4]  B Wolf,et al.  Monitoring of cellular behaviour by impedance measurements on interdigitated electrode structures. , 1997, Biosensors & bioelectronics.

[5]  Artur Dybko,et al.  Long-term three-dimensional cell culture and anticancer drug activity evaluation in a microfluidic chip. , 2013, Biosensors & bioelectronics.

[6]  T. Lee,et al.  Doxorubicin-induced apoptosis and chemosensitivity in hepatoma cell lines , 2001, Cancer Chemotherapy and Pharmacology.

[7]  P. Moela,et al.  Silencing RBBP6 (Retinoblastoma Binding Protein 6) sensitises breast cancer cells MCF7 to staurosporine and camptothecin-induced cell death. , 2014, Immunobiology.

[8]  Vítor M Gaspar,et al.  Optimization of liquid overlay technique to formulate heterogenic 3D co‐cultures models , 2014, Biotechnology and bioengineering.

[9]  C. V. van Blitterswijk,et al.  Spheroid culture as a tool for creating 3D complex tissues. , 2013, Trends in biotechnology.

[10]  Chien-Chung Peng,et al.  Drug testing and flow cytometry analysis on a large number of uniform sized tumor spheroids using a microfluidic device , 2016, Scientific Reports.

[11]  L. Ailles,et al.  Metabolic Suppression of a Drug‐Resistant Subpopulation in Cancer Spheroid Cells , 2016, Journal of cellular biochemistry.

[12]  Shuichi Takayama,et al.  Formation of stable small cell number three-dimensional ovarian cancer spheroids using hanging drop arrays for preclinical drug sensitivity assays. , 2015, Gynecologic oncology.

[13]  F. Pampaloni,et al.  The third dimension bridges the gap between cell culture and live tissue , 2007, Nature Reviews Molecular Cell Biology.

[14]  Xia Lou,et al.  High-Throughput Cancer Cell Sphere Formation for Characterizing the Efficacy of Photo Dynamic Therapy in 3D Cell Cultures , 2015, Scientific Reports.

[15]  High-Throughput Cancer Cell Sphere Formation for 3D Cell Culture. , 2017, Methods in molecular biology.

[16]  Hwan-You Chang,et al.  Recent advances in three‐dimensional multicellular spheroid culture for biomedical research , 2008, Biotechnology journal.

[17]  Tianlin Liu,et al.  Generation of Multicellular Tumor Spheroids with Microwell-Based Agarose Scaffolds for Drug Testing , 2015, PloS one.

[18]  A. Abbott Cell culture: Biology's new dimension , 2003, Nature.

[19]  P. Kiely,et al.  Using real-time impedance-based assays to monitor the effects of fibroblast-derived media on the adhesion, proliferation, migration and invasion of colon cancer cells , 2014, Bioscience reports.

[20]  Kin Fong Lei,et al.  Real-time and non-invasive impedimetric monitoring of cell proliferation and chemosensitivity in a perfusion 3D cell culture microfluidic chip. , 2014, Biosensors & bioelectronics.

[21]  Kin Fong Lei,et al.  Impedimetric quantification of the formation process and the chemosensitivity of cancer cell colonies suspended in 3D environment. , 2015, Biosensors & bioelectronics.

[22]  I. Giaever,et al.  Monitoring fibroblast behavior in tissue culture with an applied electric field. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[23]  Vladimir P Torchilin,et al.  Cancer cell spheroids as a model to evaluate chemotherapy protocols , 2012, Cancer biology & therapy.

[24]  Xiaohui Xu,et al.  Membrane Type 1 Matrix Metalloproteinase induces an epithelial to mesenchymal transition and cancer stem cell-like properties in SCC9 cells , 2013, BMC Cancer.

[25]  N. Kotov,et al.  Three-dimensional cell culture matrices: state of the art. , 2008, Tissue engineering. Part B, Reviews.

[26]  Kin Fong Lei,et al.  Quantitative Study of Cell Invasion Process under Extracellular Stimulation of Cytokine in a Microfluidic Device , 2016, Scientific Reports.

[27]  Michael Lübbert,et al.  The AML1/ETO target gene LAT2 interferes with differentiation of normal hematopoietic precursor cells. , 2014, Leukemia research.

[28]  B. Escher,et al.  Hanging drop: an in vitro air toxic exposure model using human lung cells in 2D and 3D structures. , 2013, Journal of hazardous materials.

[29]  I-Chi Lee,et al.  Label-free selection and enrichment of liver cancer stem cells by surface niches build up with polyelectrolyte multilayer films. , 2015, Colloids and surfaces. B, Biointerfaces.

[30]  Jing Liu,et al.  Soft fibrin gels promote selection and growth of tumourigenic cells , 2012, Nature Materials.

[31]  Ho-June Lee,et al.  Drug resistance via feedback activation of Stat3 in oncogene-addicted cancer cells. , 2014, Cancer cell.

[32]  Kin Fong Lei,et al.  Review on Impedance Detection of Cellular Responses in Micro/Nano Environment , 2014, Micromachines.