Tumor dissociation of highly viable cell suspensions for single-cell omic analyses in mouse models of breast cancer

Laura Rodriguez de la Fuente; Andrew M K Law; David Gallego-Ortega; Fatima Valdes-Mora

doi:10.1016/j.xpro.2021.100841

Tumor dissociation of highly viable cell suspensions for single-cell omic analyses in mouse models of breast cancer

STAR Protoc. 2021 Sep 17;2(4):100841. doi: 10.1016/j.xpro.2021.100841. eCollection 2021 Dec 17.

Authors

Laura Rodriguez de la Fuente^{1

2

3}, Andrew M K Law^{1

2}, David Gallego-Ortega^{1

2

4}, Fatima Valdes-Mora^{3

5}

Affiliations

¹ Tumour Development Group, The Kinghorn Cancer Centre, Garvan Institute of Medical Research, Sydney, NSW 2010, Australia.
² St. Vincent's Clinical School, Faculty of Medicine, University of New South Wales Sydney, Sydney, NSW 2010, Australia.
³ Cancer Epigenetic Biology and Therapeutics Lab, Children's Cancer Institute, Lowy Cancer Research Centre, University of New South Wales, Sydney, NSW 2052, Australia.
⁴ School of Biomedical Engineering, Faculty of Engineering and IT, University of Technology Sydney, Ultimo, NSW 2007 Australia.
⁵ School of Women's and Children's Health, Faculty of Medicine, University of New South Wales, Sydney, NSW 2052, Australia.

Abstract

Cell preparation with a high rate of viable cells is required to obtain reliable single-cell transcriptomic and epigenomic data. This protocol describes a technique for digestion and single-cell isolation from mouse mammary tumors to achieve ∼90% of viable cells, which can be subsequently processed in a diverse array of high-throughput single-cell "omic platforms," both in an unbiased manner or after selection of a specific cell population. For complete details on the use and execution of this protocol, please refer to Valdes-Mora et al. (2021).

Keywords: Cancer; Cell isolation; Cell separation/fractionation; Flow Cytometry/Mass Cytometry; Genomics; Model Organisms; Single Cell.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Breast Neoplasms* / genetics
Cell Separation / methods
Disease Models, Animal
Female
Humans
Mice
Single-Cell Analysis / methods
Suspensions

Substances

Suspensions