Protocol to culture and image pancreatic ductal adenocarcinoma tumor slices to study T cell migration

Nelson J. Rodríguez-Merced, Mackenzie K. Callaway, Paolo P. Provenzano

Research output: Contribution to journalArticlepeer-review

Abstract

Here, we describe a protocol for culture and live cell imaging of tumor slices. This approach studies carcinoma and immune cell dynamics in complex tumor microenvironments (TME) with nonlinear optical imaging platforms. Using a tumor-bearing mouse model of pancreatic ductal adenocarcinoma (PDA), we detail steps to isolate, activate, and label CD8+ T lymphocytes and later introduce them to live murine PDA tumor slice explants. The techniques described in this protocol can improve our understanding of cell migration in complex microenvironments ex vivo. For complete details on the use and execution of this protocol, please refer to Tabdanov et al. (2021).1

Original languageEnglish (US)
Article number102135
JournalSTAR Protocols
Volume4
Issue number1
DOIs
StatePublished - Mar 17 2023

Bibliographical note

Funding Information:
This work was supported by the NIH ( R01CA245550 , U54CA210190 University of Minnesota Physical Sciences in Oncology Center Project 2 to P.P.P., U54CA268069 CCBIR Center for Multiparametric Imaging of Tumor Immune Microenvironments RTB1 to P.P.P., and P01CA254849 Project 2 to P.P.P.). This work was also supported by an American Cancer Society Mission Boost ( MBGI-22-009-01-MBG ) award and the Randy Shaver Research and Community Fund (both to P.P.P.). We thank Hongrong Zhang and Guhan Qian for helpful comments and assistance with photos of the microscope setup.

Publisher Copyright:
© 2023 The Author(s)

Keywords

  • Cancer
  • Cell Biology
  • Immunology
  • Microscopy

PubMed: MeSH publication types

  • Journal Article
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

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