Monitoring cell fate in 3D organotypic human squamous epithelial cultures

Joshua Hatterschide; Christopher A Natale; Todd W Ridky; Elizabeth A White

doi:10.1016/j.xpro.2023.102101

Monitoring cell fate in 3D organotypic human squamous epithelial cultures

STAR Protoc. 2023 Mar 17;4(1):102101. doi: 10.1016/j.xpro.2023.102101. Epub 2023 Feb 6.

Authors

Joshua Hatterschide¹, Christopher A Natale², Todd W Ridky², Elizabeth A White³

Affiliations

¹ Department of Otorhinolaryngology: Head and Neck Surgery, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USA.
² Department of Dermatology, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USA.
³ Department of Otorhinolaryngology: Head and Neck Surgery, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USA. Electronic address: eawhite@pennmedicine.upenn.edu.

Abstract

Here, we provide a protocol to model the effects of changes to a small number of cells, such as those arising from a mutation or a virus infection, in stratified epithelia. We describe steps for diluting engineered human keratinocytes into a larger population of unmodified cells and using these cells to grow three-dimensional organotypic cultures. We detail steps to observe effects that are not apparent in homogenous organotypic epithelial cultures by visualizing the localization of modified keratinocytes in epithelial layers. For complete details on the use and execution of this protocol, please refer to Hatterschide et al. (2022).¹.

Keywords: Cancer; Cell Biology; Cell Culture; Cell Differentiation; Microbiology; Organoids; Tissue Engineering.

Publication types

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

MeSH terms

Carcinoma, Squamous Cell*
Cells, Cultured
Epithelial Cells
Epithelium
Humans
Keratinocytes*

Abstract

Publication types

MeSH terms

Grants and funding