Single Cell Secretome Analyses of Hepatic Stellate Cells: Aiming for Single Cell Phenomics

Richell Booijink, Leon Terstappen, Ruchi Bansal*

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapterAcademicpeer-review

Abstract

Activated hepatic stellate cells (HSCs) that secrete large amounts of extracellular matrix (ECM) proteins, primarily collagens, are recognized as the key pathogenic cells in liver diseases. Excessive ECM accumulation results in tissue scarring, referred to as liver fibrosis, that progresses to liver cirrhosis (liver dysfunction) and hepatocellular carcinoma. Recent studies using single cell RNA sequencing have discovered various subpopulations of HSCs with high degree of heterogeneity in quiescent, activated, as well as inactive (identified during disease regression) HSCs. However, little is known about the role of these subpopulations in ECM secretion and cell-cell communication or if they respond differently to different exogenous and endogenous factors. Moreover, how the heterogenous single cell transcriptome translates into the single cell secretome and “communicatome” (cell-cell communication) remains largely underexplored. In this chapter, we describe the method (modified enzyme-linked immunosorbent spot, ELISpot) for analyzing collagen type 1 secretion of HSCs at the single cell level, enabling a deeper understanding into the HSC secretome. In the near future, we aim to develop an integrated platform with which we can study secretome of individual cells identified by immunostaining-based fluorescence-activated cell sorting derived from healthy and diseased liver. Through the use of the VyCAP 6400-microwell chip in combination with their puncher device, we aim to perform single cell phenomics by analyzing and correlating phenotype, secretome, transcriptome, and genome of the single cells.

Original languageEnglish
Title of host publicationHepatic Stellate Cells
Subtitle of host publicationMethods and Protocols
EditorsRalf Weiskirchen, Scott L. Friedman
PublisherHumana Press
Pages257-268
Number of pages12
ISBN (Electronic)978-1-0716-3207-9
ISBN (Print)978-1-0716-3206-2978-1-0716-3206-2978-1-0716-3206-2
DOIs
Publication statusPublished - 2023

Publication series

NameMethods in Molecular Biology
Volume2669
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Collagen type 1
  • ELISpot
  • Hepatic stellate cells
  • Image analysis
  • Single cell secretome
  • NLA

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