STAR Protoc 2020 Sep 18; 1(2):100077
CA034196,JAX Director’s Innovation Fund, the British Heart Foundation, and the Leducq Foundation Transatlantic Network of Excellence in Cardiac Research.
Interstitial cells have a crucial role in cardiac fibrosis and repair of the mammalian heart. Single-cell profiling using droplet-based technology has revolutionized the investigation of cell states and identities. Here, we present a protocol for the efficient isolation of high-quality live nucleated non-cardiomyocytes from adult murine heart, for unbiased single-cell RNA sequencing using 10× Chromium technology. This protocol has been applied to homeostatic and injured hearts from different mouse strains. For complete details on the use and execution of this protocol, please refer to Forte et al. (2020).
We thank Dr. Dhanushi Abeygunawardena and Dr. Vaibhao Janbandhu for helping with the optimization of the original cell isolation protocol. We acknowledge the use of JAX Flow Cytometry, Microscopy, and Single Cell Sequencing Cores. This is an open access article under the CC BY-NC-ND license.