✍🏼 Alexsandra Zimmer, Eric R. Wang, Gaurav Choudhary, Peng Zhang

🏠 Vascular Research Laboratory, Providence VA Medical Center, Providence, RI  USA

📑 STAR Protocols (2024)

Abstract
Isolating high-quality different cell types is a powerful approach for understanding cellular compositions and features in the heart, but it is challenging. The available protocols typically focus on isolating one or two cell types. Here, we present a protocol to simultaneously isolate high-quality and high-quantity cardiomyocytes and non-myocyte cells, including immune cells, from adult rat hearts. We describe steps for purifying cells using bovine serum albumin. We also detail procedures for viability analysis and cell type identification using fluorescence-activated cell sorting.

How the WOLF was used in this study
The WOLF  was used to purify specific cell populations after tissue dissociation. Following enzymatic digestion and preparation of single-cell suspensions from adult rat hearts, the protocol uses flow cytometry—likely involving the WOLF’s microfluidic sorting capabilities—to identify and separate cardiomyocytes and non-myocyte subpopulations (including immune cells) based on fluorescent labeling and scatter properties as part of the purification process. The gentle, low-pressure microfluidic sorting provided by the WOLF sorter helps ensure high viability and integrity of isolated cells for downstream functional analyses and quality assessment of the cellular fractions obtained from the cardiac tissue.