HSCs/MPPs-Cycle, cycling hematopoietic stem cells/multipotent progenitors MEMPs, MK-Ery-mast progenitors MEMPs-Cycle, cycling MEMPs GPs, granulocytic progenitors LMPs, Ly-My progenitors pDCs, plasmacytoid dendritic cells pDCs-Cycle, cycling pDCs.ĭifferentiation Trajectory of Human Fetal Hematopoietic Cells (A) Force-directed graph (FDG) visualization of the differentiation trajectory of hematopoietic cells from Figure 1D. (D) UMAP visualization of hematopoietic cells from liver and bone marrow, colored by cell type. The colors correspond to the identified cell types. (C) Donut plots showing the percentage of transcriptionally defined (i.e., manually curated) cell populations in each of the phenotypically defined stem and progenitor populations. The standardized expression level is indicated by color intensity. For each gene, the minimum value is subtracted, and the result is divided by the maximum. The expression of the genes is standardized between 0 and 1. (B) Heatmap of the mean expression value of two manually selected marker genes for each cell type. From each fetus (17–22 pcw), phenotypically defined HSPCs and mature blood cells were sorted from bone marrow (femur and hip) and liver and processed for scRNA-seq (n = 15), scATAC-seq (n = 3), and single-cell in vitro differentiation assays (n = 4). Single-Cell Transcriptome Analysis of Human Fetal Hematopoiesis (A) Schematic overview of the experimental workflow. Our study provides a useful framework for future investigation of human developmental hematopoiesis in the context of blood pathologies and regenerative medicine.īone marrow fetal hematopoiesis fetal liver hematopoietic stem cells scATAC-seq scRNA-seq.Ĭopyright © 2020 The Authors. Finally, we refined and functionally validated the sorting strategy for the HSCs/MPPs and achieved around 90% enrichment. Integrative analysis of chromatin accessibility and gene expression revealed extensive epigenetic but not transcriptional priming of HSCs/MPPs prior to their lineage commitment. Along this trajectory, we observed opposing patterns of chromatin accessibility and differentiation that coincided with dynamic changes in the activity of distinct lineage-specific transcription factors. We inferred their differentiation trajectory and identified three highly proliferative oligopotent progenitor populations downstream of hematopoietic stem cells (HSCs)/multipotent progenitors (MPPs). Here we applied single-cell RNA sequencing (scRNA-seq) and single-cell assay for transposase-accessible chromatin sequencing (scATAC-seq) to over 8,000 human immunophenotypic blood cells from fetal liver and bone marrow. Electronic address: of hematopoiesis during human development remains poorly defined. 6 University of Cambridge, Department of Haematology, Cambridge CB2 0AW, UK Wellcome Trust - Medical Research Council Cambridge Stem Cell Institute, Cambridge CB2 0AW, UK Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton CB10 1SA, UK.5 Wellcome Trust - Medical Research Council Cambridge Stem Cell Institute, Cambridge CB2 0AW, UK.4 University of Cambridge, Department of Haematology, Cambridge CB2 0AW, UK Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton CB10 1SA, UK.3 Wellcome Trust - Medical Research Council Cambridge Stem Cell Institute, Cambridge CB2 0AW, UK Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton CB10 1SA, UK.2 European Molecular Biology Laboratory, Structural and Computational Biology Unit, Meyerhofstrasse 1, 69115 Heidelberg, Germany.1 University of Cambridge, Department of Haematology, Cambridge CB2 0AW, UK Wellcome Trust - Medical Research Council Cambridge Stem Cell Institute, Cambridge CB2 0AW, UK Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton CB10 1SA, UK.Mouse.hide() ĪttachMovie('crosshair', 'crosshair', 1) ĪttachMovie('tank', 'tank', 2, ) We need to remove bullets when they are off the stage. Obviously, if you fire a large amount of bullets, the movieclip starts lagging. Try to imagine thousands of bullets that continue falling down at increasing speed. I mean, once the bullet movieclip is on the stage, there isn’t any routine to check if the bullet should exist or not.

In the last example, when you shot a bullet, it “lives” forever. I recommend you to read the 1st part if you haven’t done it yet and we are ready to start. Welcome to the 2nd part of this tutorial.

Step seq. kongregate