Journal: Stem Cell Research & Therapy

Article Title: SARS-CoV-2-infected hiPSC-derived cardiomyocytes reveal dynamic changes in the COVID-19 hearts

doi: 10.1186/s13287-023-03603-1

Figure Lengend Snippet: Establishment of the in vitro hiPSC-derived cardiomyocyte infection system. A Workflow of proteomics analysis of SARS-CoV-2-infected heart tissues and hiPSC-derived cardiomyocytes. The proteomic data of the COVID-19 patient heart tissues were obtained from a previous study and were presented in supplementary Figure S1. After the induction of the SARS-CoV-2 infection, the hiPSC-derived cardiomyocytes’ proteome was analyzed by LC–MS/MS. B Workflow of the hiPSC-derived cardiomyocyte-based time series dynamic monitoring; hiPSC-derived cardiomyocytes were stained positive for TnT (red), α -actinin (green), and DAPI (blue) in order to distinguish the differentiated cardiomyocytes from iPSC (scale bar: 60 μm). C Immunofluorescence of the viral NP and of the cardiomyocyte marker (TnT) in SARS-CoV-2-infected hiPSC-derived cardiomyocytes at 72 h (scale bar: 50 μm)

Article Snippet: Subsequently, the medium in each well was replaced with STEMdiff Cardiomyocyte Maintenance Medium (2 mL; 05015; STEMCELL Technologies) every 2 days, for 4 times.

Techniques: In Vitro, Derivative Assay, Infection, Liquid Chromatography with Mass Spectroscopy, Staining, Immunofluorescence, Marker

Journal: Stem Cell Research & Therapy

Article Title: SARS-CoV-2-infected hiPSC-derived cardiomyocytes reveal dynamic changes in the COVID-19 hearts

doi: 10.1186/s13287-023-03603-1

Figure Lengend Snippet: Functional annotation of the proteins found to be specifically over-expressed in the SARS-CoV-2-infected hiPSC-derived cardiomyocytes at different timepoints. A According to the assessed timepoints of the over-expression protein enrichment, four patterns were identified as elevated separately at 0 (marked as yellow), 24 (light green), 48 (blue), and 72 (purple) h after the induction of the SARS-CoV-2 infection; the functional annotation of the corresponding proteins was revealed (marked separately by corresponding colors); differences at an one-way ANOVA p < 0.05 were regarded as statistically significant in the specific region; red and blue boxes indicate proteins with increased and decreased abundance, respectively. B Protein expression levels of HAND1, CCN1, NDUFA5, ATP5ME, GSTM1, SMPD1, GSTP1, MED30, NDUFB11, FIS1, XPC, BRD8, and CYFIP2 in the DIA identification groups according to the normalized protein intensity; *, p < 0.05; **, p < 0.01; ***, p < 0.001

Article Snippet: Subsequently, the medium in each well was replaced with STEMdiff Cardiomyocyte Maintenance Medium (2 mL; 05015; STEMCELL Technologies) every 2 days, for 4 times.

Techniques: Functional Assay, Infection, Derivative Assay, Over Expression, Protein Enrichment, Expressing

Journal: Stem Cell Research & Therapy

Article Title: SARS-CoV-2-infected hiPSC-derived cardiomyocytes reveal dynamic changes in the COVID-19 hearts

doi: 10.1186/s13287-023-03603-1

Figure Lengend Snippet: Functional annotation of the proteins specifically under-expressed in the SARS-CoV-2-infected hiPSC-derived cardiomyocytes at different timepoints. According to the assessed timepoints of the under-expression protein enrichment, four patterns were identified as elevated separately at 0 (marked as yellow), 24 (light green), 48 (blue), and 72 (purple) h after the induction of the SARS-CoV-2 infection; the functional annotation of the corresponding proteins was revealed (marked separately by corresponding colors); differences at an one-way ANOVA p < 0.05 were regarded as statistically significant in the specific region; red and blue boxes indicate proteins with increased and decreased abundance, respectively

Article Snippet: Subsequently, the medium in each well was replaced with STEMdiff Cardiomyocyte Maintenance Medium (2 mL; 05015; STEMCELL Technologies) every 2 days, for 4 times.

Techniques: Functional Assay, Infection, Derivative Assay, Expressing, Protein Enrichment

Journal: Stem Cell Research & Therapy

Article Title: SARS-CoV-2-infected hiPSC-derived cardiomyocytes reveal dynamic changes in the COVID-19 hearts

doi: 10.1186/s13287-023-03603-1

Figure Lengend Snippet: Co-expression profiles analysis of proteins identified the SARS-CoV-2-infected hiPSC-derived cardiomyocytes at different timepoint. A Co-expression analysis revealed six modules according to the obtained time series protein expression profiles; red and blue boxes indicate proteins with increased and decreased abundance, respectively. B , C The circos plot presents gradually over- (B, module 1 in A) and under- (C, module 4 in A) expressed proteins that are involved in multiple biological processes; terms with different colors represent different functional annotations

Article Snippet: Subsequently, the medium in each well was replaced with STEMdiff Cardiomyocyte Maintenance Medium (2 mL; 05015; STEMCELL Technologies) every 2 days, for 4 times.

Techniques: Expressing, Infection, Derivative Assay, Functional Assay

Journal: Stem Cell Research & Therapy

Article Title: SARS-CoV-2-infected hiPSC-derived cardiomyocytes reveal dynamic changes in the COVID-19 hearts

doi: 10.1186/s13287-023-03603-1

Figure Lengend Snippet: Key molecular events involved in the identified virus-host interactions. A Interaction network between DEPs and SARS-CoV-2 proteins; a map of the functional categories was used for the primary biological process analyses; yellow diamonds represent the structural, non-structural, and additional open reading frame proteins of SARS-CoV-2 extracted and identified from the cells, the gray lines represent the interactions between the viral proteins and DEPs, and the red lines represent the interactions reported in the previous study [ – ]. Immunofluorescence of the ATP energy metabolism related proteins ATP5PO and NDUFA5 B , the ROS function related proteins PTEN and CAT C , and the cardiac conduction function related proteins BIN1 and TMP1 D , combined with the cardiomyocyte marker (Actinin or TNNT2) in cardiomyocytes of COVID-19 hearts and normal controls (Scale bar: 50 µm)

Article Snippet: Subsequently, the medium in each well was replaced with STEMdiff Cardiomyocyte Maintenance Medium (2 mL; 05015; STEMCELL Technologies) every 2 days, for 4 times.

Techniques: Virus, Functional Assay, Immunofluorescence, Marker

Journal: Stem Cell Research & Therapy

Article Title: SARS-CoV-2-infected hiPSC-derived cardiomyocytes reveal dynamic changes in the COVID-19 hearts

doi: 10.1186/s13287-023-03603-1

Figure Lengend Snippet: The dynamic model of the hiPSC-derived cardiomyocytes’ proteome reveals functional changes during a SARS-CoV-2 infection. According to the dynamics of the protein expression abundance under certain temporal conditions, the DEPs can be divided into gradually and specifically DEPs, and the functional enrichment of the different patterns observed has been analyzed separately; the colors of the protein nodes indicate the measured log 2 fold-change of the proteins expressed in SARS-CoV-2-infected and control hiPSC-derived cardiomyocytes; red and blue boxes indicate proteins with up- and downregulated abundance, respectively

Article Snippet: Subsequently, the medium in each well was replaced with STEMdiff Cardiomyocyte Maintenance Medium (2 mL; 05015; STEMCELL Technologies) every 2 days, for 4 times.

Techniques: Derivative Assay, Functional Assay, Infection, Expressing, Control

Journal: bioRxiv

Article Title: Optimization of gene knockout approaches and practical solutions to sgRNA selection challenges in hPSCs with inducible Cas9 system

doi: 10.1101/2024.07.09.602644

Figure Lengend Snippet: (A) ICE analysis of Sanger sequencing chromatograms for gene knockout on each exon. (B) Significantly reduced mitochondrial potential and activity of TAZ-KO cardiomyocytes. hPSCs-CMs were stained with MitoTracker Green and imaged with a confocal microscope. Left: Representative images, scale bar = 50 μm. Right: Quantitative analysis of mitochondrial staining intensity. Two-tailed t -test.

Article Snippet: Upon reaching 90% confluency, cells were treated with cardiomyocytes differentiation basic medium (2 mM L-glutamine RPMI-1640 plus 1x B27 minus insulin) containing 5 μM CHIR99021 (STEMCELL Technologies, Canada) for 48 hours, followed by basic medium containing 5 μM IWR-1-endo (STEMCELL Technologies) for another 48 h. The cells were then cultured in basic medium for more than 6 days until spontaneous contractions were observed.

Techniques: Sequencing, Gene Knockout, Activity Assay, Staining, Microscopy, Two Tailed Test