Project description:Tridimensional cardiac differentiation from hiPSCs has been largely described in the literature. However, the exact impact that 3D culture has throughout the entire process of cardiac differentiation remains poorly defined. We developed a robust and efficient 3D platform for cardiomyocyte differentiation from hiPSCs, based on the temporal modulation of WNT signalling using small molecules. 3D aggregates of hiPSCs were generated by forced aggregation in microwells and subsequently differentiated. In order to determine the differences in gene expression profile due to 3D culture throughout the different stages of cardiac differentiation, we compared transcriptional changes between cells in 3D aggregates and standard 2D monolayer cardiac differentiation. Analysis of these data suggests a faster commitment of hiPSCs toward the cardiac lineage and also higher degree of cardiomyocyte functional maturation after 20 days of culture in the 3D aggregates when compared with the 2D monolayer.
Project description:In vitro cardiac differentiation can be readily achieved in human induced pluripotent stem cells (hiPSCs) via temporal modulation of Wnt signalling. However, its differentiation trajectory and the in vivo counterpart of intermediate progenitor populations has yet to be elucidated. This study aims to capture the most diverse amount of distinct cell populations during in vitro cardiac differentiation by performing single cell RNA-sequencing at day 2, 4, 5 and 9 cells undergoing in vitro cardiac differentiation. Our study has revealed in vitro cardiac differentiation is analogous with in vivo second heart field cell specification.
Project description:We performed miRNA array analysis to analyze the miRNAs secreted into the supernatant during the mesoderm/cardiac differentiation and maturation process from human induced pluripotent stem cells (hiPSCs). Supernatant samples were collected on day -1, day 3, day 5, day 7, day 9, day 21, day 35, and day 51.
Project description:We reported loss of HBL1 lncRNA promoted cardiogenesis. Here we wanted to know the genomewide gene expression profiles caused by HBL1 KO during cardiac differentiation.
Project description:In this study, isotretinoin (INN)-induced alternations in transcriptome during caidiomyocyte differentiation derived from human hESCs and hiPSCs were investigated. H1-hESC and C15-hiPSC were differentiated to caidiomyocytes under exposure to sublethal level of INN, and cells were collected at day 0 (undifferentiated cellsl) day 2 (mesoderm) and day 6 (cardiac progenitors) for genome-wide transcriptomic profiling by RNA-seq.
Project description:The role of FGF-MEK-ERK signalling pathway during embryonic heart development has not been fully elucidated. Here, we inhibited the pathway for 1 day using PD0325901, a MEK inhibitor, at the lateral plate mesoderm stage during cardiac differentiation of human embryonic stem cells. Cells were collected on day 2 (before PD0325901 administration), day 3 and day 8 to determine the effect of a transient FGF-MEK-ERK pathway modulation on the cardiac cell fate choice.
Project description:We show that a synthetic modified messenger RNA (smRNA)-based reprogramming method that leads to the generation of transgene-free OLs has been developed. An smRNA encoding a modified form of OLIG2, a key TF in OL development, in which the serine 147 phosphorylation site is replaced with alanine, OLIG2S147A, is designed to reprogram hiPSCs into OLs. We demonstrate that repeated administration of the smRNA encoding OLIG2 S147A lead to higher and more stable protein expression. Using the single-mutant OLIG2 smRNA morphogen, we establish a 6-day smRNA transfection protocol, and glial induction lead to rapid NG2+ OL progenitor cell (OPC) generation (> 70% purity) from hiPSC-derived neural progenitor cells (NPCs). The smRNA-induced NG2+ OPCs can mature into functional OLs in vitro and promote remyelination in vivo. Proteomic analysis of OLIG2-binding proteins indicates that OLIG2 is bound by the heat shock protein 70 (HSP70) complex. The HSP70 complex is bound more strongly to OLIG2 with the modified phosphorylation site than to wild-type OLIG2.