Cell Biology, Faculty of Medicine and Health Technology, Tampere University, FI-33014 Tampere, Finland.
FICAM, Faculty of Medicine and Health Technology, Tampere University, FI-33014 Tampere, Finland.
Int J Mol Sci. 2020 Mar 6;21(5):1833. doi: 10.3390/ijms21051833.
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.
药物治疗旨在抑制肿瘤血管生成，以阻止肿瘤生长。然而，并非所有肿瘤类型都对这种治疗方法有反应。一种能够识别诱导血管生成的癌症类型的筛选方法将有助于预测抗血管生成药物对患者的疗效。我们的目标是开发（1）一种细胞检测方法，以评估患者来源的肿瘤细胞的血管生成诱导潜力，和（2）一种在血管平台上培养癌细胞的方案。我们优化了培养基组成和细胞（hASC、HUVEC 和癌细胞）的接种密度，以适应 48 孔、96 孔甚至 384 孔板的尺寸，以允许血管形成和癌细胞增殖，并随后进行高通量分析。通过定量分析管状结构的形成，研究了患者来源的癌细胞的血管生成诱导潜力，并通过基因表达和细胞活力（WST-1 测定）评估了在血管平台上生长的癌细胞对药物的反应。免疫细胞化学用血管性血友病因子、胶原 IV、CD44、细胞角蛋白 19 和 ALDH1A1 进行。血管生成诱导潜力测试对癌细胞诱导的血管生成具有反应性。当在血管平台或塑料上生长时，癌细胞的反应在基因表达水平和活力结果上有所不同。这两个方案是为个性化医学选择有效癌症药物和作为替代癌细胞培养平台提供有希望的新工具。
