Generating cell-derived matrices from human trabecular meshwork cell cultures for mechanistic studies

Felix Yemanyi, Janice Vranka, Vijay Krishna Raghunathan

Research output: Chapter in Book/Report/Conference proceedingChapter

4 Scopus citations

Abstract

Ocular hypertension has been attributed to increased resistance to aqueous outflow often as a result of changes in trabecular meshwork (TM) extracellular matrix (ECM) using in vivo animal models (for example, by genetic manipulation) and ex vivo anterior segment perfusion organ cultures. These are, however, complex and difficult in dissecting molecular mechanisms and interactions. In vitro approaches to mimic the underlying substrate exist by manipulating either ECM topography, mechanics, or chemistry. These models best investigate the role of individual ECM protein(s) and/or substrate property, and thus do not recapitulate the multifactorial extracellular microenvironment; hence, mitigating its physiological relevance for mechanistic studies. Cell-derived matrices (CDMs), however, are capable of presenting a 3D-microenvironment rich in topography, chemistry, and whose mechanics can be tuned to better represent the network of native ECM constituents in vivo. Critically, the composition of CDMs may also be fine-tuned by addition of small molecules or relevant bioactive factors to mimic homeostasis or pathology. Here, we first provide a streamlined protocol for generating CDMs from TM cell cultures from normal or glaucomatous donor tissues. Second, we document how TM cells can be pharmacologically manipulated to obtain glucocorticoid-induced CDMs and how generated pristine CDMs can be manipulated with reagents like genipin. Finally, we summarize how CDMs may be used in mechanistic studies and discuss their probable application in future TM regenerative studies.

Original languageEnglish (US)
Title of host publicationMethods in Cell Biology
EditorsDavid Caballero, Subhas C. Kundu, Rui Luís Reis
PublisherAcademic Press Inc.
Pages271-307
Number of pages37
ISBN (Print)9780128201725
DOIs
StatePublished - 2020

Publication series

NameMethods in Cell Biology
Volume156
ISSN (Print)0091-679X

Keywords

  • Cell-derived matrix
  • Extracellular matrix
  • Glaucoma
  • Mechanobiology
  • Trabecular meshwork

ASJC Scopus subject areas

  • Cell Biology

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