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Protocol for Isolation of Cardiac Interstitial Cells from Adult Murine Hearts for Unbiased Single Cell Profiling.

STAR protocols (2020-10-02)
Elvira Forte, Sandra Daigle, Nadia A Rosenthal
ABSTRACT

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).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
牛血清白蛋白 来源于牛血清, lyophilized powder, ≥96% (agarose gel electrophoresis)
Sigma-Aldrich
肝素 钠盐 来源于猪肠粘膜, Grade I-A, ≥180 USP units/mg
Sigma-Aldrich
红细胞裂解缓冲液Hybri-Max, liquid, sterile-filtered, suitable for hybridoma
Sigma-Aldrich
碘化丙啶 溶液, solution (1.0 mg/ml in water)