Figure 2.

Lentiviral tagging and HTS demonstrates trilineage mesoderm differentiation of monoclonal hPSC. (A): Schematic of monoclonal hPSC lineage tagging. A single transduced hPSC with 25 lentiviral tags was clonally expanded, induced to form embryonic mesoderm progenitors (hEMP) and differentiated into H, E, and M lineages, which were then isolated as separate populations by FACS and subjected to vector integration site analysis by HTS. X is the set of all 25 lentiviral tags found in the initial hPSC clone. (B): Table listing all 25 lentiviral tags detected by HTS in the hPSC clone and the three lineages differentiated from hEMP generated from the hPSC clone, with chromosome, strand and position information. Checkmarks indicate that a particular tag was detected in the lineage indicated at the top of the column. Each row in schematic heatmap to the right represents a distinct lentiviral tag, equivalent to the rows of the table on left. A filled in box indicates the presence of that tag in the population, and the three populations (E, H, and M) are annotated as green, red and blue colors respectively for clarity. Numbers in parentheses under each lineage label refer to the total number of tags found in each population, and numbers on the y-axis indicate the total number of tags found among all populations. All 25 lentiviral tags were detected as shared among the three lineages, as indicated in the table to the right of the heatmap. Abbreviations: E, endothelial; H, hematopoietic; hEMP, human embryonic mesoderm progenitor; hPSC, human pluripotent stem cell; M, mesenchymal.