Assessing the heterogeneity of cardiac non-myocytes and the effect of cell culture with integrative single cell analysis

Cardiac non-myocytes comprise a diverse and crucial cell population in the heart that plays dynamic roles in cardiac wound healing and growth. Non-myocytes broadly fall into four cell types: endothelium, fibroblasts, leukocytes, and pericytes. Here we characterize the diversity of the non-myocytes i...

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Published inbioRxiv
Main Authors Iskra, Brian, Davis, Logan, Miller, Henry E, Yu-Chiao, Chiu, Bishop, Alexander R, Chen, Yidong, Aune, Gregory J
Format Paper
LanguageEnglish
Published Cold Spring Harbor Cold Spring Harbor Laboratory Press 05.03.2020
Subjects
CD11b antigen
CD200 antigen
CD45 antigen
CD9 antigen
Cell culture
Cell proliferation
Cytometry
Endothelium
Fibroblasts
Heart
Leukocytes
Myeloid cells
Myocytes
Neural networks
Pericytes
Ribonucleic acid
RNA
Surface markers
Wound healing
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Summary:Cardiac non-myocytes comprise a diverse and crucial cell population in the heart that plays dynamic roles in cardiac wound healing and growth. Non-myocytes broadly fall into four cell types: endothelium, fibroblasts, leukocytes, and pericytes. Here we characterize the diversity of the non-myocytes in vivo and in vitro using mass cytometry. By leveraging single-cell RNA sequencing we inform the design of a mass cytometry panel. To aid in annotation of the mass cytometry datasets, we utilize data integration with a neural network. We introduce approximately 460,000~ single cell proteomes of non-myocytes as well as 5,000~ CD31 negative single cell transcriptomes. Using our data, as well as previously reported datasets, we characterize cardiac non-myocytes with high depth in six mice, characterizing novel surface markers (CD9, CD200, Notch3, and FolR2). Further, we find that extended cell culture promotes the proliferation of CD45+CD11b+FolR2+IAIE- myeloid cells in addition to fibroblasts.
DOI:10.1101/2020.03.04.975177