A 3D Heterotypic Multicellular Tumor Spheroid Assay Platform to Discriminate Drug Effects on Stroma versus Cancer Cells

Weydert, Zoe; Mathews-Greiner, Lesley; Thiel, Christoph; Cordes, Henrik; Küpfer, Lars; Guye, Patrick; Kelm, Jens M.; Ferrer, Marc

doi:10.1177/2472555219880194

Cited by 21 publications

(18 citation statements)

References 30 publications

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“…6,7 During the past decade, 3D spheroid models have also been adapted for use in assay-and automation-ready plate formats, including 96-, 384-, and 1536-well plates, as well as emerging microfluidic systems that support organ networks. [8][9][10][11][12][13] With minor modifications, 3D spheroid models can be easily adapted to validated cellular endpoints, including assays originally designed for 2D cell culture to assess cell viability, cell function, and gene expression (e.g., biochemical assays, secreted biomarker assays, and omics methods). 14,15 Cell-based assays, however, often rely on lytic and secretion endpoints and interrogate the whole spheroid, not specific cell populations or spatially defined regions of the spheroid.…”

Section: Introductionmentioning

confidence: 99%

A Framework for Optimizing High-Content Imaging of 3D Models for Drug Discovery

et al. 2020

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Section: Introductionmentioning

confidence: 99%

A Framework for Optimizing High-Content Imaging of 3D Models for Drug Discovery

et al. 2020

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“…3D models provide enormous benefit beyond conventional 2D cell culture models with regards to establishing an efficient in vitro pre-clinical platform for drug screening that accurately represents the original tumor [10, 19, 42]. Several 3D models have been developed including organoids, tissue slices, hydrogels, bioreactors, microfluidic models and scaffold-free spheroids [10–14, 16–18, 35].…”

Section: Discussionmentioning

confidence: 99%

Patient-derived tumor spheroid cultures as a promising tool to assist personalized therapeutic decisions in breast cancer

Hofmann¹,

Cohen-Harazi²,

Maizels³

et al. 2021

Preprint

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Breast cancer is the most common cause of cancer related death in women. Treatment of breast cancer has many limitations including a lack of accurate biomarkers to predict success of chemotherapy and intrinsic resistance of a significant group of patients to the gold standard of therapy. Therefore, new tools are needed to provide doctors with guidance in choosing the most effective treatment plan for a particular patient and thus to increase the survival rate for breast cancer patients. Here, we present a successful method to grow in vitro spheroids from primary breast cancer tissue. Samples were received in accordance with relevant ethical guidelines and regulations. After tissue dissociation, in vitro spheroids were generated in a scaffold-free 96-well plate format. Spheroid composition was investigated by immunohistochemistry (IHC) of epithelial (Pan Cytokeratin (panCK)), stromal (Vimentin) and breast cancer-specific markers (ER, PR, HER2, GATA-3). Growth and cell viability of the spheroids were assessed upon treatment with multiple anti-cancer compounds. Students t-test and two-way ANOVA test were used to determine statistical significance. We were able to successfully grow spheroids from 27 out of 31 samples from surgical resections of breast cancer tissue from previously untreated patients. Recapitulation of the histopathology of the tissue of origin was confirmed. Furthermore, a drug panel of standard first- and second-line chemotherapy drugs used to treat breast cancer was applied to assess the viability of the patient-derived spheroids and revealed variation between samples in the response of the spheroids to different drug treatments. We investigated the feasibility and the utility of an in vitro patient-derived spheroid model for breast cancer therapy, and we conclude that spheroids serve as a highly effective platform to explore cancer therapeutics and personalized treatment efficacy. These results have significant implications for the application of this model in clinical personalized medicine.

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“…27–29 Heterotypic or complex spheroids can also be grown from multiple cell types. 30–36 The incorporation of stromal cells into tumor models has been shown to affect the proliferation, intracellular signaling, and gene expression of the malignant cells. 25,33,37,38 Including stromal cells in the tumor spheroid model increased malignant cell survival and resistance to anticancer agents, 30–32 which may reflect more physiologically relevant pharmacological responses.…”

Section: Introductionmentioning

confidence: 99%

“…30–36 The incorporation of stromal cells into tumor models has been shown to affect the proliferation, intracellular signaling, and gene expression of the malignant cells. 25,33,37,38 Including stromal cells in the tumor spheroid model increased malignant cell survival and resistance to anticancer agents, 30–32 which may reflect more physiologically relevant pharmacological responses. Here we describe the development, optimization, and validation of a complex tumor spheroid assay, incorporating both human malignant and stromal cell components, for HTS.…”

Section: Introductionmentioning

confidence: 99%

Complex Tumor Spheroids, a Tissue-Mimicking Tumor Model, for Drug Discovery and Precision Medicine

et al. 2021

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Malignant tumors are complex tissues composed of malignant cells, vascular cells, structural mesenchymal cells including pericytes and carcinoma-associated fibroblasts, infiltrating immune cells, and others, collectively called the tumor stroma. The number of stromal cells in a tumor is often much greater than the number of malignant cells. The physical associations among all these cell types are critical to tumor growth, survival, and response to therapy. Most cell-based screens for cancer drug discovery and precision medicine validation use malignant cells in isolation as monolayers, embedded in a matrix, or as spheroids in suspension. Medium- and high-throughput screening with multiple cell lines requires a scalable, reproducible, robust cell-based assay. Complex spheroids include malignant cells and two normal cell types, human umbilical vein endothelial cells and highly plastic mesenchymal stem cells, which rapidly adapt to the malignant cell microenvironment. The patient-derived pancreatic adenocarcinoma cell line, K24384-001-R, was used to explore complex spheroid structure and response to anticancer agents in a 96-well format. We describe the development of the complex spheroid assay as well as the growth and structure of complex spheroids over time. Subsequently, we demonstrate successful assay miniaturization to a 384-well format and robust performance in a high-throughput screen. Implementation of the complex spheroid assay was further demonstrated with 10 well-established pancreatic cell lines. By incorporating both human stromal and tumor components, complex spheroids might provide an improved model for tumor response in vivo.

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A 3D Heterotypic Multicellular Tumor Spheroid Assay Platform to Discriminate Drug Effects on Stroma versus Cancer Cells

Cited by 21 publications

References 30 publications

A Framework for Optimizing High-Content Imaging of 3D Models for Drug Discovery

A Framework for Optimizing High-Content Imaging of 3D Models for Drug Discovery

Patient-derived tumor spheroid cultures as a promising tool to assist personalized therapeutic decisions in breast cancer

Complex Tumor Spheroids, a Tissue-Mimicking Tumor Model, for Drug Discovery and Precision Medicine

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