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In Vitro Vascular Network Modified to Function as Culture Platform and Angiogenic Induction Potential Test for Cancer
Outi Huttala1,2,3, Synnöve Staff3,4, Tuula Heinonen2
1Cell Biology, Faculty of Medicine and Health Technology, Tampere University, FI-33014 Tampere, Finland.
Abstract:
Drug treatments have been designed to inhibit tumor angiogenesis in hope of stopping tumor growth. However, not all tumor types respond to this type of treatment. A screening method which identifies angiogenesis inducing cancer types would help predict the efficacy of angiogenesis-inhibiting drugs for the patients. Our goal is to develop (1) a cell assay to assess the angiogenic induction potential of patient-derived tumor cells, and (2) a protocol for culturing cancer cells on a vascular platform. We optimized the media composition and seeding density of cells (hASC, HUVEC, and cancer cells) to 48-, 96-, and even 384-well plate sizes to allow vascular formation and cancer cell proliferation and subsequent analysis with high throughput. The angiogenic induction potential of patient-derived cancer cells was investigated by quantifying the formation of tubular structures and the drug response of cancer cells grown on a vascular platform was evaluated using gene expression and cell viability (WST-1) assay. Immunocytochemistry was performed with von Willebrand factor, collagen IV, CD44, cytokeratin 19 and ALDH1A1. The angiogenic induction potential test was shown to be responsive to the induction of angiogenesis by cancer cells. The responses of cancer cells were different when grown on a vascular platform or on plastic, seen in gene expression level and viability results. These two protocols are promising novel tools for aiding the selection of efficient cancer drugs for personalized medicine and as an alternative cancer cell culture platform.
Insights
Developing novel cell assays to assess tumor angiogenesis potential and drug response. This aids in predicting treatment efficacy for personalized cancer medicine.
Area of Science:
- Oncology
- Biotechnology
- Cell Biology
Background:
- Tumor angiogenesis is a target for cancer drug development, but efficacy varies by tumor type.
- Predictive assays are needed to identify patients likely to benefit from anti-angiogenic therapies.
Purpose of the Study:
- To develop a cell assay for assessing angiogenic potential of patient-derived tumor cells.
- To establish a vascular platform for culturing cancer cells and evaluating drug responses.
Main Methods:
- Optimized cell culture conditions (media, seeding density) in multi-well plates (48- to 384-well).
- Assessed angiogenic induction by quantifying tubular structure formation.
- Evaluated drug response using gene expression and cell viability (WST-1) assays on vascular platforms.
- Performed immunocytochemistry for specific biomarkers (vWF, collagen IV, CD44, CK19, ALDH1A1).
Main Results:
- The developed assay effectively detected cancer cell-induced angiogenesis.
- Cancer cells exhibited different gene expression and viability responses on vascular platforms compared to plastic.
- The angiogenic induction potential test showed responsiveness to cancer cell activity.
Conclusions:
- Developed high-throughput cell assay and vascular platform protocols for cancer research.
- These tools can aid in selecting effective cancer drugs for personalized medicine.
- The vascular platform serves as an alternative to traditional plastic culture for cancer cells.
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