In Vitro Vascular Network Modified to Function as Culture Platform and Angiogenic Induction Potential Test for Cancer Cells

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.

Decellularized In Vitro Capillaries for Studies of Metastatic Tendency and Selection of Treatment

“…Because DCs do not have living cells, DC experiments can be designed according to the requirements of the cancer cells, and parameters such as culture time can be adjusted according to the needs of the cells in question. The scalability of the vascular network [ 22 ] also allows the use of DC in high-throughput studies.…”

“…Isolation of cancer cells was performed as described earlier [ 22 ]. Briefly, liquid patient samples (pleural effusion or ascites fluid) were centrifuged at 200× g for 10 min, and cell pellets were washed, strained through 100 µm strainer, and resuspended to Liquid cancer sample medium (LCM, Table 1 ).…”

“…The use of the tissue samples and correlating data was approved by Ethics Committee of the Pirkanmaa Hospital District, Tampere, Finland, with permit number R16127 (dated 7 February 2017). Isolation of cancer cells was performed as described earlier [22]. Briefly, liquid patient samples (pleural effusion or ascites fluid) were centrifuged at 200× g for 10 min, and cell pellets were washed, strained through 100 µm strainer, and resuspended to Liquid cancer sample medium (LCM, Table 1).…”

“…We have previously developed human cell-based in vitro capillaries that can be used as a growth platform for primary cancer cell cultures [ 22 ]. These capillaries, which form a network of capillaries in culture, contain a lumen, an endothelial cell layer surrounded by contractile cells, and a basement membrane, including relevant attachment proteins found in in vivo vessels [ 23 ].…”