In Vitro Culture and Histological Evaluation of 3D Organotypic Cultures

William Dalleywater; Francesca Wheat; Declan Sculthorpe; Georgina Hyland; Mohammad Ilyas

doi:10.1007/978-1-0716-3076-1_13

In Vitro Culture and Histological Evaluation of 3D Organotypic Cultures

Methods Mol Biol. 2023:2650:155-170. doi: 10.1007/978-1-0716-3076-1_13.

Authors

William Dalleywater^{1

2}, Francesca Wheat³, Declan Sculthorpe⁴, Georgina Hyland⁴, Mohammad Ilyas^{4

5}

Affiliations

¹ Translational Medical Sciences, School of Medicine, University of Nottingham, Nottingham, UK. William.dalleywater@nottingham.ac.uk.
² Department of Cellular Pathology, Nottingham University Hospitals NHS Trust, Nottingham, UK. William.dalleywater@nottingham.ac.uk.
³ Department of Cellular Pathology, University Hospitals of Leicester NHS Trust, Nottingham, UK.
⁴ Translational Medical Sciences, School of Medicine, University of Nottingham, Nottingham, UK.
⁵ Department of Cellular Pathology, Nottingham University Hospitals NHS Trust, Nottingham, UK.

PMID: 37310631
DOI: 10.1007/978-1-0716-3076-1_13

Abstract

Organotypic cultures allow cells to grow in a system which mimics in vivo tissue organization. Here we describe a method for establishing 3D organotypic cultures (using intestine as an example system), followed by methods for demonstrating cell morphology and tissue architecture using histological techniques and molecular expression analysis using immunohistochemistry, though the system is also amenable to molecular expression analysis, such as by PCR, RNA sequencing, or FISH.

Keywords: 3D organotypic cultures; Immunohistochemistry; In vitro; Intestine.

MeSH terms

Histological Techniques*
Microphysiological Systems*
Polymerase Chain Reaction