Project description:Human pluripotent stem cells (hPSC) offer a unique cellular model to study the molecular details underpinning embryonic development. We profiled single-cell RNA-seq (scRNA-seq) on 957 single cells collected at 6 time points over a 30-day period of non-directed differentiation of human neural progenitor cells (NPC) generated from hESC. Computational and experimental analyses of this comprehensive data set enabled us to: 1) define distinct subpopulations of developing neurons and their specific gene expression patters; 2) elucidate alternative cell developmental trajectories within discrete time points of the differentiating populations; and 3) decipher gene regulatory networks and validated key regulators (transcription factors and lincRNAs) that dictate alternative cell fate specifications. In this submission, 957 bam files are provided: 1) 573 single cells from time course profiling (day 0, 1, 5, 7, 10, 30) as first batch and 2) 384 single cells from time course profiling (day 0, 3, 7, 14) as second batch.
Project description:H3K4me3 ChIP-Seq profiling of a directed differentiation time course converting human embryonic stem cells (hES) into immature pancreatic beta cell precursors. CyT49, a Viacyte proprietary male hESC line with normal karyotype was used.
Project description:Aims: Pluripotent stem cell-derived endothelial cell products possess therapeutic potential in ischemic vascular disease. However, the factors that drive endothelial differentiation from pluripotency and cellular specification are largely unknown. The aims of this study were to use single cell RNA sequencing (scRNA-seq) to map the transcriptional landscape and cellular dynamics of directed differentiation of human embryonic stem cell-derived endothelial cells (hESC-EC), and to compare these cells to mature endothelial cells from diverse vascular beds.Methods and Results: A highly efficient directed 8-day differentiation protocol was used to generate a hESC-derived endothelial cell product (hESC-ECP), in which 66% of cells coexpressed CD31 and CD144. We observed largely homogeneous hESC and mesodermal populations at days 0 and 4 respectively, followed by a rapid emergence of distinct endothelial and mesenchymal populations. Pseudotime trajectory identified transcriptional signatures of endothelial commitment and maturation during the differentiation process. Concordance in transcriptional signatures was verified by scRNA-seq analysis using both a second hESC line RC11, and an alternative hESC-EC differentiation protocol. In total 105,727 cells were subjected to scRNA-seq analysis. Global transcriptional comparison revealed a transcriptional architecture of hESC-EC that differs from freshly isolated and cultured human endothelial cells and from organ-specific endothelial cells. Conclusions: A transcriptional bifurcation into endothelial and mesenchymal lineages was identified, as well as novel transcriptional signatures underpinning commitment and maturation. The transcriptional architecture of hESC-ECP was distinct from mature and foetal human EC.
Project description:To assess changes in the miRNA-ome during the early stages of endothelial differentiation, the expression of mature human miRNAs at days 2, 4 and 10 of endothelial differentiation (vs pluripotent time-matched samples) in SA461 hESC cell line was analysed in a two-channel microarray. Expression of the pluripotency-associated miRNAs miR-302a – d and miR-372 and miR-373 was significantly suppressed with progression of differentiation. The expression of miR-99b, miR-181a and miR-181b was, in contrast, increased in a time- and differentiation-dependent manner compared to time-matched pluripotent samples. The expression profiles were validated and good concordance was observed between microarray, Q-PCR and Northern blot analysis. This experiment includes a total of 28 samples which are divided in to 3 groups, each with 4 biological repeats: group 1, Parental pluripotent hESC control sample. group 2, time-matched pluripotent hESC control samples. Group 3, hESC cells directed toward endothelial differentiation at time points 2 days, 4 days and 10 days.
Project description:Efficient directed cardiac induction of human embryonic stem cells requires an intermediate inhibition of WNT signaling, shortly after the initial induction of mesoderm. To reveal the significance of the WNT inhibition step during differentiation, two differential time-course analyses were performed - both comparing WNT-inhibited and non-WNT-inhibited samples: In the first time-course, the WNT inhibitor IWP-2 was either added or omitted on days 2 and 3, and in the second time-course, IWP-2 was either added or omitted on days 1 and 2.
Project description:In order to provide multi-omic resolution to human retinal organoid developmental dynamics, we performed scRNA-seq and scATAC-seq from the same cell suspension across a time course (6-46 weeks) of human retinal organoid development. This data set covers all the retinal organoid scRNA-seq data generated from IMR90 and409B2-iCas9 cell lines.
Project description:To understand the function of CHD8 in human ESC and neural differentiation, we generated the CHD8 knockout human ESCs and characterize the effect of loss-of-function of CHD8 on pluripotency maintenance and neuroectoderm determination by utilizing an efficient neuroectoderm differentiation protocol. Samples from hESC, CHD8KO_hESC and their derived-neural progenitor cells (hNPC) were collected for high-throughput sequencing with at least two replicates for each sample. Differentially expressed genes analysis showed loss of CHD8 affect the genes in ESC and NPC.
Project description:Transcriptional profiling of a directed differentiation time course converting human embryonic stem cells (hES) into immature pancreatic beta cell precursors.
