Protocol for culturing patient-derived organoids of cervical cancer

Rui Wang; Timothy Harris; Dalissa Negrón-Figueroa; David Lo; Allison Judge; D'Shaunique Walters; Bo Jiang; Lauren E Colbert

doi:10.1016/j.xpro.2024.103353

Protocol for culturing patient-derived organoids of cervical cancer

STAR Protoc. 2024 Oct 1;5(4):103353. doi: 10.1016/j.xpro.2024.103353. Online ahead of print.

Authors

Rui Wang¹, Timothy Harris¹, Dalissa Negrón-Figueroa¹, David Lo¹, Allison Judge¹, D'Shaunique Walters², Bo Jiang³, Lauren E Colbert⁴

Affiliations

¹ Department of Radiation Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA.
² Department of Radiation Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA; The University of Texas MD Anderson Cancer Center UTHealth Houston Graduate School of Biomedical Sciences, Houston, TX 77030, USA.
³ Department of Thoracic/Head and Neck Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA.
⁴ Department of Radiation Oncology, The University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA. Electronic address: lcolbert@mdanderson.org.

Abstract

Herein, we present a protocol for culturing patient-derived organoids (PDOs) of cervical cancer that includes workflows for tumor biopsy/resection tissue and cytobrush-sampled cells. We describe steps for PDO culture initiation, including rinsing, gentle dissociation, Lymphoprep separation, and cell assessment, as well as seeding cells from surgical and cytobrush tissue digestion. We then provide guidance on PDO maintenance and passage and techniques for producing conditioned medium. Overall, this protocol serves as a valuable guide for establishing and maintaining cervical cancer PDOs. For complete details on the use and execution of this protocol, please refer to Colbert et al.¹.

Keywords: cancer; genomics; metabolism; organoids.