Live-Cell Sender-Receiver Co-cultures for Quantitative Measurement of Paracrine Signaling Dynamics, Gene Expression, and Drug Response

Michael Pargett; Abhineet R Ram; Vaibhav Murthy; Alexander E Davies

doi:10.1007/978-1-0716-3008-2_13

Live-Cell Sender-Receiver Co-cultures for Quantitative Measurement of Paracrine Signaling Dynamics, Gene Expression, and Drug Response

Methods Mol Biol. 2023:2634:285-314. doi: 10.1007/978-1-0716-3008-2_13.

Authors

Michael Pargett¹, Abhineet R Ram¹, Vaibhav Murthy^{2

3}, Alexander E Davies^{4

5}

Affiliations

¹ Department of Molecular and Cellular Biology, University of California, Davis, Davis, CA, USA.
² Department of Veterinary Biosciences, College of Veterinary Medicine, The Ohio State University, Columbus, OH, USA.
³ Knight Cancer Institute, Cancer Early Detection Advanced Research Center, Oregon Health & Science University, Portland, OR, USA.
⁴ Department of Veterinary Biosciences, College of Veterinary Medicine, The Ohio State University, Columbus, OH, USA. daviesal@ohsu.edu.
⁵ Knight Cancer Institute, Cancer Early Detection Advanced Research Center, Oregon Health & Science University, Portland, OR, USA. daviesal@ohsu.edu.

PMID: 37074584
DOI: 10.1007/978-1-0716-3008-2_13

Abstract

Paracrine signaling is a fundamental process regulating tissue development, repair, and pathogenesis of diseases such as cancer. Herein we describe a method for quantitatively measuring paracrine signaling dynamics, and resultant gene expression changes, in living cells using genetically encoded signaling reporters and fluorescently tagged gene loci. We discuss considerations for selecting paracrine "sender-receiver" cell pairs, appropriate reporters, the use of this system to ask diverse experimental questions and screen drugs blocking intracellular communication, data collection, and the use of computational approaches to model and interpret these experiments.

Keywords: Cancer; Drug screening; ERK; Intercellular communication; Live-cell microscopy; MAPK; Single cell.

Publication types

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

MeSH terms

Cell Culture Techniques
Coculture Techniques
Gene Expression
Paracrine Communication*
Signal Transduction*

Abstract

Publication types

MeSH terms

Grants and funding