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

Yichen Guo; Bronte Miller; Michael Heim; Ana Gutierrez-Garcia; Renata Jaskula-Sztul; Bin Ren; Mary Kathryn Sewell-Loftin

doi:10.1016/j.xpro.2023.102177

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

STAR Protoc. 2023 Apr 21;4(2):102177. doi: 10.1016/j.xpro.2023.102177. Online ahead of print.

Authors

Yichen Guo¹, Bronte Miller², Michael Heim², Ana Gutierrez-Garcia³, Renata Jaskula-Sztul⁴, Bin Ren⁵, Mary Kathryn Sewell-Loftin⁶

Affiliations

¹ Department of Biomedical Engineering, University of Alabama at Birmingham, Birmingham AL 35294, USA; Department of Surgery, University of Alabama at Birmingham, Birmingham AL 35233, USA.
² Department of Biomedical Engineering, University of Alabama at Birmingham, Birmingham AL 35294, USA.
³ Department of Surgery, University of Alabama at Birmingham, Birmingham AL 35233, USA.
⁴ Department of Surgery, University of Alabama at Birmingham, Birmingham AL 35233, USA; O'Neal Comprehensive Cancer Center, University of Alabama at Birmingham, Birmingham AL 35233, USA.
⁵ Department of Biomedical Engineering, University of Alabama at Birmingham, Birmingham AL 35294, USA; Department of Surgery, University of Alabama at Birmingham, Birmingham AL 35233, USA; O'Neal Comprehensive Cancer Center, University of Alabama at Birmingham, Birmingham AL 35233, USA. Electronic address: bren@uabmc.edu.
⁶ Department of Biomedical Engineering, University of Alabama at Birmingham, Birmingham AL 35294, USA; O'Neal Comprehensive Cancer Center, University of Alabama at Birmingham, Birmingham AL 35233, USA. Electronic address: mksewellloftin@uab.edu.

Abstract

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.

Keywords: Biotechnology and Bioengineering; Cancer; Cell Biology; Cell Culture; Cell-based Assays; Microscopy; Molecular Biology; Tissue Engineering.

Abstract

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