Protocol for preparing monosodium urate crystals for in vitro and in vivo studies in mouse and human cells

Mohnish Alishala; Ru Liu-Bryan; Isidoro Cobo

doi:10.1016/j.xpro.2024.103030

Protocol for preparing monosodium urate crystals for in vitro and in vivo studies in mouse and human cells

STAR Protoc. 2024 Apr 27;5(2):103030. doi: 10.1016/j.xpro.2024.103030. Online ahead of print.

Authors

Mohnish Alishala¹, Ru Liu-Bryan², Isidoro Cobo³

Affiliations

¹ Department of Cellular and Molecular Medicine, University of California San Diego, 9500 Gilman Drive, La Jolla, CA 92093, USA.
² Division of Rheumatology, Allergy, and Immunology, UCSD School of Medicine, 9500 Gilman Drive, La Jolla, CA 92093, USA.
³ Division of Clinical Immunology & Rheumatology, Department of Medicine, Heersink School of Medicine, 1825 University Blvd, Birmingham, AL 35294, USA. Electronic address: isidorocobo@uabmc.edu.

Abstract

Gout is caused by the deposition of monosodium urate crystals (MSUc) in the joints, triggering a unique inflammatory and metabolic response in macrophages. Here, we present a protocol to generate MSUc for in vitro and in vivo studies in mouse and human cells. We describe steps for dissolving uric acid followed by crystallizing, purifying, evaluating, and analyzing MSUc. We then detail procedures for stimulating human/mouse-derived macrophages and determining endotoxin levels in MSUc preparation.

Keywords: Cell Biology; Molecular Biology.