h9 human embryonic stem cells hescs  (WiCell Research Institute Inc)

Journal: Stem Cell Research & Therapy

Article Title: FHOD3 deficiency disrupts sarcomere organization and activates caMKII signaling in human stem cell-derived cardiomyocytes

doi: 10.1186/s13287-026-04902-z

Figure Lengend Snippet: Generation of FHOD3-deficient hESC model. A Diagram of the FHOD3 knockout pattern, showing the gene editing position and deleted base pair. B Immunofluorescence staining of the human pluripotency markers SSEA4 and OCT4 in WT and FHOD3 KO hESCs. Scale bar, 75 μm. C QPCR analysis of SSEA4 and OCT4 expression in WT and FHOD3 KO hESCs, n = 3 per group. D Diagram of cardiomyocytes differentiation from hESCs by using small molecule-based methods. E Verification of FHDO3 knockout by WB, Full-length blots are presented in Supplementary Figure S3. F Flow cytometry and quantification of the myocardial-specific marker cardiac Troponin T (TNNT2) in WT and FHOD3 KO cardiomyocytes after 15days of differentiation, n = 3 per group. Data are represented as mean ± SEM; * P < 0.05, ** P < 0.01, *** P < 0.001. Data were analyzed via two-sample t test followed by the Bonferroni post hoc test

Article Snippet: Human embryonic stem cell (hESC) line H9 (WA09, WiCell) was used in this study.

Techniques: Knock-Out, Immunofluorescence, Staining, Expressing, Flow Cytometry, Marker

Journal: Stem Cell Research & Therapy

Article Title: FHOD3 deficiency disrupts sarcomere organization and activates caMKII signaling in human stem cell-derived cardiomyocytes

doi: 10.1186/s13287-026-04902-z

Figure Lengend Snippet: FHOD3 deficiency results in cardiomyocytes abnormality and sarcomere disassembly. A Immunofluorescence staining of sarcomeric proteins cTNT and α-actinin to showed significant larger percentage of disrupted and disorganized sarcomeres in FHOD3 KO hESC-CMs (30 days of differentiation). Scale bar, 10 μm. B Transmission electron microscopy images of sarcomere structures in WT and FHOD3 KO hESC-CMs (30 days of differentiation) and quantification of abnormal sarcomeres. Scale bar, 100 nm. C , D Calibration of forward scatter (FSC) in flow cytometry (FSC;10000 cells/ sample) to shows the volume of cardiomyocytes (30 days of differentiation), n = 3 per group. E , F Immunofluorescence of phalloidin in WT and FHOD3 KO hESC-CMs (30 days of differentiation) to show the mean cell area, Scale bar, 50 μm. Data are represented as mean ± SEM, * P < 0.05, ** P < 0.01, *** P < 0.001. Data were analyzed via two-sample t test followed by the Bonferroni post hoc test

Article Snippet: Human embryonic stem cell (hESC) line H9 (WA09, WiCell) was used in this study.

Techniques: Immunofluorescence, Staining, Transmission Assay, Electron Microscopy, Flow Cytometry

Journal: Stem Cell Research & Therapy

Article Title: FHOD3 deficiency disrupts sarcomere organization and activates caMKII signaling in human stem cell-derived cardiomyocytes

doi: 10.1186/s13287-026-04902-z

Figure Lengend Snippet: FHOD3 knockout cardiomyocytes exhibit compromised contractile functions. A Schematic diagram of contractility measurement in WT and FHOD3 KO hESC-CMs (30 days of differentiation). B The curve of contraction versus time detected by ‘MUSCLEMOTION’. C Quantification of contraction amplitude, time to peak, relaxation time and 90% contraction duration, n = 6 per group. D Representative Ca 2+ transient signals in WT and FHOD3 KO hESC-CMs (30 days of differentiation) measured by using Fluo-4 AM. E Waveform diagram of calcium transient. F Schematic diagram of calcium transient measurement indicators. G , H Ca 2+ transient induced by 10 mmol/L caffeine and waveform diagram. I Quantification of amplitude, time to peak value and 50% decay time of calcium transient in (D), n = 10 per group. J Quantification of amplitude, time to peak value and 50% decay time of calcium transient induced by 10 mmol/L caffeine in (G), n = 4 per group. Data are represented as mean ± SEM; * P < 0.05, ** P < 0.01, *** P < 0.001. Data were analyzed via two-sample t test followed by the Bonferroni post hoc test

Article Snippet: Human embryonic stem cell (hESC) line H9 (WA09, WiCell) was used in this study.

Techniques: Knock-Out

Journal: Stem Cell Research & Therapy

Article Title: FHOD3 deficiency disrupts sarcomere organization and activates caMKII signaling in human stem cell-derived cardiomyocytes

doi: 10.1186/s13287-026-04902-z

Figure Lengend Snippet: Different transcriptome between WT and KO hESC-CMs (30 days of differentiation). A The volcano plot shows differentially expressed genes between two groups ( n = 3, respectively). Red denotes up-regulated genes, whereas green represents down-regulated genes. B KEGG Enrichment Scatter Plot dispalys the top 20 significant pathways. C KEGG Enrichment Chord Diagram displays the 10 Genes with the highest fold change (left) in the top 9 significant pathways (right). D The scatter plot of GO enrichment for differentially expressed genes. E QPCR analysis of the genes involved sarcomere structure and calcium handling pathways, normalized to IPO8, n = 3 per group

Article Snippet: Human embryonic stem cell (hESC) line H9 (WA09, WiCell) was used in this study.

Techniques:

Journal: Stem Cell Research & Therapy

Article Title: FHOD3 deficiency disrupts sarcomere organization and activates caMKII signaling in human stem cell-derived cardiomyocytes

doi: 10.1186/s13287-026-04902-z

Figure Lengend Snippet: FHOD3 knockout hESC-CMs develop mitochondrial dysfunction. A Seahorse Mito Stress Test evaluating mitochondiral respiration in WT and KO hESC-CMs (30 days of differentiation), Left: real-time OCR profiles; Right: quantification of mitochondrial respiration parameters, n = 6 per group. B ATP production detection in WT and KO hESC-CMs (30 days of differentiation), n = 6 per group. C Representative fluorescence images of ROS levels in WT and KO hESC-CMs (30 days of differentiation) and quantification analysis, n = 7 per group. Enzymatic activity of individual ETC complexes I, IV and V assessed by using commercial assay kits (30 days of differentiation), n = 6 per group. Data are represented as mean ± SEM; * P < 0.05, ** P < 0.01, *** P < 0.001. Data were analyzed via two-sample t test followed by the Bonferroni post hoc test

Article Snippet: Human embryonic stem cell (hESC) line H9 (WA09, WiCell) was used in this study.

Techniques: Knock-Out, Fluorescence, Activity Assay

Journal: Stem Cell Research & Therapy

Article Title: FHOD3 deficiency disrupts sarcomere organization and activates caMKII signaling in human stem cell-derived cardiomyocytes

doi: 10.1186/s13287-026-04902-z

Figure Lengend Snippet: FHOD3 deficiency activates calcium signaling pathway to promote heart failure progression. A Representative Western blot showing sarcomere structure proteins in WT and FHOD3 KO hESC-CMs (30 days of differentiation), Quantification shown below, Full-length blots are presented in Supplementary Figure S5. B Representative Western blot showing calcium related proteins in WT and FHOD3 KO hESC-CMs (30 days of differentiation), Quantification shown below, Full-length blots are presented in Supplementary Figure S6. C QPCR analysis of cardiac remodeling markers reglated by the CAMKII downstream effectors in WT and FHOD3 KO hESC-CMs (30 days of differentiation), n = 3 per group.Data are represented as mean ± SEM; * P < 0.05, ** P < 0.01, *** P < 0.001. Data were analyzed via two-sample t test followed by the Bonferroni post hoc test

Article Snippet: Human embryonic stem cell (hESC) line H9 (WA09, WiCell) was used in this study.

Techniques: Western Blot

Journal: Stem Cell Research & Therapy

Article Title: FHOD3 deficiency disrupts sarcomere organization and activates caMKII signaling in human stem cell-derived cardiomyocytes

doi: 10.1186/s13287-026-04902-z

Figure Lengend Snippet: Candidate myosin activator OM rescues myocardial contractile dysfunction caused by FHOD3 deficiency. A The curve of contraction versus time detected by ‘MUSCLEMOTION’. B Quantification of contraction amplitude, time to peak, relaxation time and 90% contraction duration in WT and FHOD3 KO hESC-CMs (30 days of differentiation), n = 8 per group. C Representative Ca 2+ transient signals in WT and FHOD3 KO hESC-CMs (30 days of differentiation) measured by using Fluo-4 AM, and waveform diagram of calcium transient. D Quantification of amplitude, time to peak value and 50% decay time of calcium transient in (C), n = 10 per group. Data are represented as mean ± SEM; * P < 0.05, ** P < 0.01, *** P < 0.001. Data were analyzed via one-way ANOVA followed by the Bonferroni post hoc test

Article Snippet: Human embryonic stem cell (hESC) line H9 (WA09, WiCell) was used in this study.

Techniques:

Journal: bioRxiv

Article Title: Human ectodermal organoids reveal the cellular origin of DiGeorge Syndrome

doi: 10.1101/2025.08.08.669417

Figure Lengend Snippet: A) A heatmap showing differentially expressed genes that also include known markers for each respective differentiated NC derivative cell type at different timepoints of early and fully differentiated stages. B ) A 30-day protocol produces a pure population of pigmented MITF-immunopositive melanocytes that also express TYR and PMEL. MITF qPCR NC SD = ±0.002; Day 30 Melanocytes SD = ±4.919. MITF immuno percentage positive NC SD = 0; Day 30 Melanocytes SD = 0. TYR qPCR NC SD = ±0.008; Day 30 Melanocytes SD = ±223.7. PMEL immuno NC SD = ±0.021; Day 30 Melanocytes SD = ±0.6033. C ) Virtually all cells in the glial cultures differentiated into S100B-immunopositive cells. S100B qPCR NC SD = ±0.9851; D26 SD = ±2.476; D35 SD = ±2.853. S100B immuno NC SD = ±0.0001; D26 SD = ±0.0355; D35 SD = ±0.2323 D ) Virtually all sensory neurons express the cell type specific marker BRN3A. BRN3A qPCR NC SD = ±0.4091; D10 SD = ±20.05. BRN3A immuno D10 = ± 8.315, D16 = ±9.708. E ) Symphatetic neurons differentiated into a pure population of TH-immunopositive cells that also significantly increased expression of TBH at the end of the 16-day protocol. DBH qPCR NC SD = ±0.6501, D10 SD = ±0.7824, D16 SD = ±5.811. DBH imunno D10 SD = ±21.46, D16 SD = ±0. TH qPCR NC SD = ±0.3162, D10 SD = ±2.576, D16 SD = ±5.205. F ) Differentiation into chrommaffin cell lineage formed PNMT-positive cells in ten days. PNMT qPCR NC SD = ±0.1704, D10 SD = ±3.229. PNMT immuno Isotype SD = ±0.1115, D5 SD = ±0.1396, D10 SD = ±1.080. G ) Chondrocyte cultures formed Aggrecan and SOX9 -positive mineralizing chondrospheroids in 28 days. The cartilage lineage initiation and NC marker SOX9 expression is highest in early and midpoint cultures. SOX9 qPCR NC SD = ±0.4912, D14 SD = ±0.2429, D28 SD = ±0.07434. SOX9 immuno NC SD = ±0.1266, D14 SD = ±0.0939, D28 SD = ±0.1296. ACAN qPCR NC SD = ±0.0015, D14 SD = ±0.2845, D28 = ± 0.0596. ACAN immuno NC = ±0.0591, D14 SD = ±0.1144, D28 = ±0.1303. H ) In 28 days, NC cells organized into RUNX2-immunopositive osteocytes that have nuclei synchronized in a symmetrical angle. RUNX2 qPCR NC SD = ±0.0642, D14 SD = ±0.2367, D28 SD = ±0.3629. I ) Adipocytes express increasing levels of FABP4 during their 33 days on maturation resulting in a homogeneous culture of cells that form lipid-spot positive lipid droplets. FABP4 qPCR NC SD = ±0.0010, D11 SD = ± 0.0288, D33 SD = ±0.2864. LipidSpot immuno NC SD = ± 0.0942, D11 = ±0.1615, D33 = ±0.4709 J ) Transgelin and Smooth Muscle Actin expressing cells show a typical smooth muscle morphology after 21 days in a uniform culture. ACTA qPCR NC SD = ±1.269, D11 SD = ±2.676, D21 SD = ±5.413. ACTA immune NC SD = ±0.036, D11 SD = ±0.2735, D21 SD = ±0.4533. TAGLN qPCR NC SD = ±2.739, D11 SD = ±28.70, D21 = ±46.39. TAGLN immuno NC SD = ±0.0798, D11 SD = ±0.3547, D21 SD = ±0.5395. ISO= isotype IgG control (the images stained by additional antibodies are shown in ). Significance is annotated as such *: p-value > 0.05, **: p-value >0.002, ***: p-value >0.0002, ****: p-value >0.0001. NC: Neural Crest, Mel: Melanocytes, Sens Neu: Sensory Neurons, Symp Neu: Sympathetic Neurons, Chrom: Adrenal Chromaffin Cells, Chond: Chondrocytes, Osteo: Osteocytes, Adi: Adipocytes, Smo Mus: Smooth Muscle, hESC: human embryonic stem cells.

Article Snippet: H1 and H9 human embryonic stem cell (hESC) lines (WA09 (H9) and WA01 (H1) from WiCell, were used for the characterization of the ectodermal organoids.

Techniques: Marker, Expressing, Control, Staining