A team led by Eva van Rooij at the prestigious Hubrecht Institute in Utrecht has undertaken a significant transcriptomics study in the search for drug targets for the encouragement of cardiac tissue repair following damage due to ischemia.
Using their tissue dissociation and FACS techniques they were able to analyse single adult cardiomyocytes of mice following cardiac ischemic injury. To ensure that they sorted intact, live and nucleated cells the single cell suspension was stained with DAPI and DRAQ5 respectively.
The initial findings for cardiomyocytes, fibroblasts, neutrophils, endothelial cells and macrophages point to a significant role for beta-2 microglobulin, secreted from stressed cardiomyocytes and activating fibroblasts to stimulate repair for example.
The combination of DAPI and DRAQ5™ has become the benchmark for sorting of single cells for downstream molecular analysis, as exemplified in this work. For a detailed analysis of the rationale see this earlier blog article and Ordoñez-rueda, et al.
References:
Molenaar, Bas, et al. "Single-cell transcriptomics following ischemic injury identifies a role for B2M in cardiac repair." Communications Biology 4.1 (2021): 1-15.
Ordoñez‐Rueda, Diana, et al. "Apoptotic Cell Exclusion and Bias‐Free Single‐Cell Selection Are Important Quality Control Requirements for Successful Single‐Cell Sequencing Applications." Cytometry Part A 97.2 (2020): 156-167.
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