Characterization of iCell cardiomyocytes using single-cell RNA-sequencing methods
| Autor: | Tobias Hildebrandt, Patrick Baum, Christina Schmid, Christian T. Wohnhaas, Georg Rast |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
TNNT2
Induced Pluripotent Stem Cells Cell Population Cardiomyocytes Cell cycle hiPSC Human induced pluripotent stem cell derived cardiomyocytes iCell cardiomyocytes MYL7 P4HB Single-cell sequencing Vimentin Drug Evaluation Preclinical Cell Separation 030204 cardiovascular system & hematology Biology Toxicology 030226 pharmacology & pharmacy Cell Line 03 medical and health sciences 0302 clinical medicine ddc:570 medicine Humans Myocyte Myocytes Cardiac RNA-Seq Induced pluripotent stem cell education Fibroblast Pharmacology education.field_of_study Cell Cycle Cell Differentiation Fibroblasts Cell biology medicine.anatomical_structure Single-Cell Analysis Transcriptome Biomarkers |
| Popis: | IntroductionHuman induced pluripotent stem cell (hiPSC)-derived cardiomyocytes are being evaluated for their use in pharmacological and toxicological testing, particularly for electrophysiological side effects. However, little is known about the composition of the commercially available iCell cardiomyocyte (Fuijifilm Cellular Dynamics) cultures and the transcriptomic phenotype of individual cells.MethodsWe characterized iCell cardiomyocytes (assumed to be a mixture of nodal-, atrial-, and ventricular-like cardiomyocytes together with potential residual non-myocytes) using bulk RNA-sequencing, followed by investigation of cellular heterogeneity using two different single-cell RNA-sequencing platforms.ResultsBulk RNA-sequencing identified key cardiac markers (TNNT2, MYL7) as well as fibroblast associated genes (P4HB, VIM), and cardiac ion channels in the iCell cardiomyocyte culture. High-resolution single cell RNA-sequencing demonstrated that both, cardiac and fibroblast-related genes were co-expressed throughout the cell population. This approach resolved two cell clusters within iCell cardiomyocytes. Interestingly, these clusters could not be associated with known cardiac subtypes. However, transcripts of ion channels potentially useful as functional markers for cardiac subtypes were below the detection limits of the single-cell approaches used. Instead, one cluster (10.8% of the cells) is defined by co-expression of cardiac and cell cycle-related genes (e.g. TOP2A). Incorporation of bromodeoxyuridine further confirmed the capability of iCell cardiomyocytes to enter cell cycle.DiscussionThe co-expression of cardiac related genes with cell cycle or fibroblast related genes may be interpreted either as aberrant or as an immature feature. However, this excludes the presence of a non-cardiomyocyte sub-population and indicates that some cardiomyocytes themselves enter cell cycle. published |
| Databáze: | OpenAIRE |
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