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

doi:10.1016/j.xpro.2023.102135

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

Biomedical Engineering

Research output: Contribution to journal › Article › peer-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 language	English (US)
Article number	102135
Journal	STAR Protocols
Volume	4
Issue number	1
DOIs	https://doi.org/10.1016/j.xpro.2023.102135
State	Published - 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

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Publisher link

10.1016/j.xpro.2023.102135

Find It

Find It at U of M Libraries

Cite this

@article{792b794c4ce84f9683742738b007b776,

title = "Protocol to culture and image pancreatic ductal adenocarcinoma tumor slices to study T cell migration",

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",

keywords = "Cancer, Cell Biology, Immunology, Microscopy",

author = "Rodr{\'i}guez-Merced, {Nelson J.} and Callaway, {Mackenzie K.} and Provenzano, {Paolo P.}",

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: {\textcopyright} 2023 The Author(s)",

year = "2023",

month = mar,

day = "17",

doi = "10.1016/j.xpro.2023.102135",

language = "English (US)",

volume = "4",

journal = "STAR Protocols",

issn = "2666-1667",

publisher = "Cell Press",

number = "1",

}

TY - JOUR

T1 - Protocol to culture and image pancreatic ductal adenocarcinoma tumor slices to study T cell migration

AU - Rodríguez-Merced, Nelson J.

AU - Callaway, Mackenzie K.

AU - Provenzano, Paolo P.

N1 - 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)

PY - 2023/3/17

Y1 - 2023/3/17

N2 - 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

AB - 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

KW - Cancer

KW - Cell Biology

KW - Immunology

KW - Microscopy

UR - http://www.scopus.com/inward/record.url?scp=85148766760&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85148766760&partnerID=8YFLogxK

U2 - 10.1016/j.xpro.2023.102135

DO - 10.1016/j.xpro.2023.102135

M3 - Article

C2 - 36861840

AN - SCOPUS:85148766760

SN - 2666-1667

VL - 4

JO - STAR Protocols

JF - STAR Protocols

IS - 1

M1 - 102135

ER -

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

Abstract

Bibliographical note

Keywords

UN SDGs

Publisher link

Find It

Other files and links

Fingerprint

Cite this