Skip to main content

Research Repository

Advanced Search

Quantitative methods for measuring repair rates and innate-immune cell responses in wounded mouse skin

Li, Zhi; Gothard, Elizabeth; Coles, Mark C; Ambler, Carrie A

Quantitative methods for measuring repair rates and innate-immune cell responses in wounded mouse skin Thumbnail


Authors

Zhi Li

Elizabeth Gothard

Mark C Coles



Abstract

In skin wounds, innate immune cells clear up tissue debris and microbial contamination, and also secrete cytokines and other growth factors that impact repair process such as re-epithelialization and wound closure. After injury, there is a rapid influx and efflux of immune cells at wound sites, yet the function of each innate cell population in skin repair is still under investigation. Flow cytometry is a valuable research tool for detecting and quantifying immune cells, however in mouse back skin, the difficulty in extracting immune cells from small area of skin due to tissue complexity has made cytometric analysis an underutilized tool. Here in this paper, we provide detailed methods on the digestion of lesion-specific skin without disrupting antigen expression followed by multiplex cell staining that allows for identification of seven innate immune populations, including rare subsets such as group 3 innate lymphoid cells (ILC3s), by flow cytometry analysis. Further, when studying the functions of immune cells to tissue repair an important metric to monitor is size of the wound opening. Normal wounds close steadily albeit at non-linear rates, while slow or stalled wound closure can indicate an underlying problem with the repair process. Calliper measurements are difficult and time-consuming to obtain and can require repeated sedation of experimental animals. We provide advanced methods for measuring of wound openness; digital 3D image capture and semi-automated image processing that allows for unbiased, reliable measurements that can be taken repeatedly over time.

Citation

Li, Z., Gothard, E., Coles, M. C., & Ambler, C. A. (2018). Quantitative methods for measuring repair rates and innate-immune cell responses in wounded mouse skin. Frontiers in Immunology, 9, Article 347. https://doi.org/10.3389/fimmu.2018.00347

Journal Article Type Article
Acceptance Date Feb 7, 2018
Online Publication Date Feb 27, 2018
Publication Date Feb 27, 2018
Deposit Date Feb 7, 2018
Publicly Available Date Mar 29, 2024
Journal Frontiers in Immunology
Publisher Frontiers Media
Peer Reviewed Peer Reviewed
Volume 9
Article Number 347
DOI https://doi.org/10.3389/fimmu.2018.00347

Files

Published Journal Article (2.1 Mb)
PDF

Copyright Statement
Copyright: © 2018 Li, Gothard, Coles and Ambler. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.





You might also like



Downloadable Citations