An Automatable Hydrogel Culture Platform for Evaluating Efficacy of Antibody‐Based Therapeutics in Overcoming Chemoresistance

Abstract

The microenvironment plays a major role in conferring chemoresistance to cancer cells. In order to better inform clinical response to chemoresistance, preclinical models that recapitulate its hallmark features are needed to enable screening for resistance-specific therapeutic targets. A novel platform for seeding cancer cells in 3D hydrogels is presented utilizing derivatives of chitosan and alginate that, critically, is amenable to high throughput screening: cell seeding in hydrogels, media changes, dosing of anticancer compounds, and cell viability assays are all automated using a standard and commercially available liquid handling robot. Culture in these hydrogels elicits resistance in ovarian, lung, and prostate cancer cells to treatment by doxorubicin and paclitaxel. In correlation, proteomics analysis of SKOV3 cells cultured in 3D reveals enrichment of proteins associated with extreme drug resistance including HMOX1 and ALDH2. Subsequently, therapeutic antibodies targeted to tumor-associated antigens upregulated in 3D cultures are shown to have higher efficacy compared to 2D cultures. Collectively, this automated 3D cell culture platform provides a powerful tool with utility in identification of drugs that may overcome chemoresistance. declare no conflict of interest. C.M. is a cofounder of the spinoff company CellSpring AG, a company in which he owns shares.

Effect of Wenxia Changfu Formula Combined With Cisplatin Reversing Non-Small Cell Lung Cancer Cell Adhesion-Mediated Drug Resistance

“…It is widely used in A549 cells, MDA-MB-231 cells, melanoma cell line SK-Mel-28 cells, and prostate epithelial tumor promoter cells ( Fetah et al., 2019 ). Agarose has been widely used as a scaffold for cell culture and has also been successfully used in the 3D culture of tumor cells ( Kletzmayr et al., 2020 ). In 3D model, the architecture of cancer tumor is more complex, cell spheroids are commonly divided into 3 layers: the outer layer with high proliferation rates, the middle layer with senescent cells, and the hypoxic core with necrotic cells ( Amann et al., 2014 ; Hoarau-Véchot et al., 2018 ).…”

Polymeric Hydrogels for In Vitro 3D Ovarian Cancer Modeling

“…Kletzmayr et al [ 197 ] developed a chemically modified alginate derivative through an oxidation process (oxAlg) to form hydrogels via chemical crosslinking with N-succinyl-chitosan (sChi). The developed hydrogel, obtainable inside the 96-well plates classically used in 2D cell culture studies, was able to support the growth of ovarian, lung, and prostate cancer cells, and provided a fast and reliable new platform for high-throughput drug screening ( Figure 9 a–c).…”

Clinical Application Perspectives of Lung Cancers 3D Tumor Microenvironment Models for In Vitro Cultures

“…Blache et al used polyethylene-glycol hydrogels to form a 3D platform for simultaneous analysis of transcriptome and secretome [108]. Hydrogels also became a tool for tumor neovascularization study through the mimicking of blood vessel formation in vitro [104,[109][110][111][112][113][114][115][116][117][118]. Development of the liquid phase in hydrogels and tumor-on-chip models provided the possibility for the replacement of the culture fluid and thus ensuring the flow of cellular metabolites and regulatory molecules [88,116,119].…”