Protocol for indirect and direct co-culture between human cancer cells and endothelial cells

Abstract

Summary: The cross talk between cancer cells and endothelial cells (ECs) within the tumor microenvironment plays a critical role in tumor progression, recurrence, and cancer stemness. Here, we present a protocol containing two in vitro approaches to study such interactions. We first describe an indirect co-culture system to study the regulation of stemness markers in cancer cells by secreted factors from ECs. We then detail a direct co-culture system to study juxtracrine communications between the cell types.For complete details on the use and execution of this protocol, please refer to Sewell-Loftin et al.1 and Guo et al.2 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Keywords

• Biotechnology and Bioengineering
• Cancer
• Cell Biology
• Cell Culture
• Cell-based Assays
• Microscopy