Extended Data Fig. 2: Verification of whole organoid barcoded lineage tracing by FACS in organoids and barcoded lineage tracing in developing mouse embryos.
From: Cerebral organoids display dynamic clonal growth and tunable tissue replenishment
a, In silico subsampling of indicated percentages of lineages in organoids with 90% confidence range of subsampled results indicated. b, Scheme of fluorescent H9-based WT cells lines generated. c, Example of flow cytometry separation of fluorescent cell lines in equal mixtures. d, Cell line contribution of fluorescent cell lines in chimeric organoids mixed at 90:2:2:2:2:2 contribution of cell lines (‘Blue’, ‘Cyan’, ‘Purple’, ‘Green’, ‘Yellow’, ‘Red’) over time. (n = 3 organoids as biological replicates from 1 differentiation experiment at each time point). Data points represent mean ± SEM. e, Outline of barcoded lineage tracing in mice from E9.5, E10.5 and E13.5 to E18.5. f, In utero injection of two fluorescent retroviruses carrying GFP and RFP at E10.5 with analysis following on E17.5. One embryo hemisphere was cleared and imaged entirely for GFP and RFP. Individual z-sections are indicated. Note the lack of overlap of GFP and RFP signal indicating single viral integrations. g, Lineage size distributions in lineage traced mouse brains. Individual Barcode Libraries 1 and 2 as well as the number of detected lineages are indicated. Left and right hemispheres as well as midbrain of individual embryos were scored as indicated. (n = individual lineages from 4 mouse brains as biological replicates split into left ventricle, right ventricle and midbrain at each time point).
