hesc line h9 wa09  (WiCell Research Institute Inc)

Journal: Advanced Science

Article Title: Comprehensive Profiling of N 6 ‐methyladnosine (m 6 A) Readouts Reveals Novel m 6 A Readers That Regulate Human Embryonic Stem Cell Differentiation

doi: 10.1002/advs.202510075

Figure Lengend Snippet: Assessment of the regulatory effects of m 6 A‐binding proteins and m 6 A readers on hESC proliferation and differentiation. (A) Representative images of morphology of WT versus FUBP3 , FXR2 , and L1TD1 KO hESCs under bright field. (B) Growth curve assessing cell proliferation kinetics of FUBP3 , FXR2 , and L1TD1 KO versus WT hESCs. (C) ALP staining assay of WT versus FUBP3 , FXR2 , and L1TD1 KO hESCs. (D) Schematic representation of the induction endodermal differentiation experiment of hESCs. (E‐F) Flow cytometry and statistical histogram analysis of endodermal differentiation of WT versus FUBP3 , FXR2 and L1TD1 KO hESCs. (G) Schematic representation of the induction hematopoietic mesodermal differentiation experiment of hESCs. (H‐I) Flow cytometry and statistical histogram analysis of hematopoietic mesodermal differentiation of WT versus FUBP3 , FXR2 and L1TD1 KO hESCs. (J) Schematic representation of the induction neural ectodermal differentiation experiment of hESCs. (K‐L) Flow cytometry and statistical histogram analysis of neural ectodermal differentiation of WT versus FUBP3 , FXR2 and L1TD1 KO hESCs. (M‐O) RT‐qPCR analysis of definitive endoderm markers SOX17, FOXA2 (M); mesoderm marker TBXT (also known as T) and CD34 (N); and neural‐ectoderm markers SOX1, PAX6, NES in WT versus FUBP3 , FXR2 and L1TD1 KO hESCs (O). Figures in (D), (G), and (J) were drawn with Biovisart ( https://biovisart.com.cn ) and Bioicons ( https://bioicons.com/ ) with permission. Data in (B), (F), (I), (L), and (M‐O) were statistically analyzed using two‐tailed unpaired Student's t ‐test. Error bars represent mean ± SD ( n = 3 independent experiments). For all statistical plots, * means p < 0.05, ** means p < 0.01, *** means p < 0.001, **** means p < 0.0001, ns means not significant.

Article Snippet: The human embryonic stem cell (hESC) line H9 (WA09) was obtained from WiCell Research Institute (USA, catalog#WA09, RRID: CVCL_9773) in 2019 under appropriate material transfer agreements.

Techniques: Binding Assay, Staining, Flow Cytometry, Quantitative RT-PCR, Marker, Two Tailed Test

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: