Anne-Marie Galow

The project addresses the generation and establishment of programmed pacemaker cells for an in vitro drug testing possibility to perform predictive tests. This may lead to an improved treatment of cardiac arrhythmias or an accurate identification of potential drug molecules at a very early stage of development. Important benefits will arise in verifying the safety of a wide variety of medicines while reducing animal testing. For more information you may visit our project website at ...

Single-cell RNA-sequencing (scRNA-seq) provides high-resolution insights into complex tissues. Cardiac tissue, however, poses a major challenge due to the delicate isolation process and the large size of mature cardiomyocytes. Regardless of the experimental technique, captured cells are often impaired and some capture sites may contain multiple or no cells at all. All this refers to “low quality” potentially leading to data misinterpretation. Common standard quality control parameters involve the ...

This study contains our snRNA-Seq based comparison of whole hearts from Fzt.DU and Bl6 mice published in Cardiovascular Research.

This study includes the single snRNA-seq in whole adult murine hearts from an inbred (C57BL/6NRj) and an outbred (Fzt:DU) mouse strain in comparison to publicly available scRNA-seq data of the tabula muris project.

This study contains our single nuclei characterisation of whole hearts from Fzt.DU mice published in Cells.

For scRNA-Seq, iSABs were dissociated using the Primary Cardiomyocyte Isolation Kit (Thermo Fisher Scientific) before library preparation was performed using the 10xGenomics system with subsequent sequencing on the HighSeq4000 (Illumina). The mouse-SAN scRNA-Seq protocol is described in Goodyer et al. Preprocessing of raw sequencing data from iSABs relied on tools of the Cell Ranger Software (v.6.1.0) as was the procedure in Goodyer et al. Downstream analyses were conducted similar for both ...

This pdf file contains all figures of the article "Integrative Cluster Analysis of Whole Hearts Reveals Proliferative Cardiomyocytes in Adult Mice" in high resolution.

This pdf contains the top 10 markers for each identified cell type as a dot-plot.

The file contains an UMAP representation illustrating the expression of the stem cell marker Cd34 in all identified cell clusters.

The file contains FeaturePlots of 4 different cardiomyocyte markers (Actn2, Tnnc1, Actc1 and Ryr2) to demonstrate how annotation of clusters needs to be based on several markers and approaches, which in the aggregate allow for more reliable conclusions/results.

The file contains the absolute and relative cell/nuclei numbers per cluster for each individual dataset used in the integrated analysis. Subpopulations were grouped together for the representation of relative cell/nuclei numbers.

This file contains the top 100 transcripts for the identified cell types in the integrated dataset.

This dot-plot represents only the most significant genes per identified cell type cluster.

This file contains the top markers for the identified cell types.

This pdf contains the top 10 markers for each identified cell type as a dot-plot.

This file contains the R-script to analyse single nuclei data previously processed with kallisto and bustools. The analyses utilizes the Seurat, harmony and RNAvelocity package.

This file contains the script for the downstream scRNA-Seq analysis including quality control using the barcode ranking method together with the tool DropletUtils to exclude empty droplets and undetected genes as well as standard data processing (normalisation, variable feature identification, scaling, and dimensionality reduction) using tools of Seurat (v.3.2.2). After cluster annotation the %mtDNA was plotted for both datasets.

This file contains the detailed experimental protocol for the cultivation of "induced sinoatrial bodies (iSABs), the scRNA-Seq procedure as well as the general computational workflow for data processing. The R-script is provided separately.

This file contains the R-script to analyse single nuclei and single cell data of Bl6 and Fzt:DU mice previously processed with cellranger. The analyses utilizes the Seurat and harmony package to integrate three datasets before subsequent downstream analysis characterizing proliferative cardiomyocytes.

This file contains the detailed experimental protocol to isolate nuclei from murine hearts.

This file contains Violin plots for mitochondrial gene transcripts (%mtRNA), cardiac marker (Tnnt2), and pace-maker marker (Hcn4) for identified cell clusters in iSABS and sinoatrial node region (results obtained from a data reanalysis of Goodyer et al.).

This file contains several Scatter plots illustrating the correlation of %mtDNA and (a) Tnnt2 (b) Hcn4 (c) total number of gene transcripts per cell In Tnnt2 positive cells and (d) total number of gene transcripts per cell In Hcn4 positive cells.

Graphical abstract for the manuscript "Quality control in scRNA-Seq can discriminate pacemaker cells – the mtDNA bias" by Anne-Marie Galow#, Sophie Kussauer, Markus Wolfien, Robert David*, and Andreas Hoeflich#*

This file contains a UMAP plot generated for the Goodyer dataset illustrating the various cell populations and their proportions as well as the original t-SNE plot of the respective data published by Goodyer et al. in 2019 (doi: 10.1161/CIRCRESAHA.118.314578).

This file contains a UMAP plot for the iSABs dataset illustrating the various cell populations and their proportions.

This file contains a detailed description of the underlying experimental procedure and the computational analysis.