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Fig. 2 | Stem Cell Research & Therapy

Fig. 2

From: Genetic barcoding reveals clonal dominance in iPSC-derived mesenchymal stromal cells

Fig. 2

Generation of iMSCs is based on few dominant subclones. a Schematic presentation of the experimental workflow with lentiviral transduction (LV-trans.) of iPSCs. After puromycin selection, the genetic labelling with RGB-BC was either analyzed during long-term culture of iPSCs, or upon differentiation towards iMSCs. bd Sequencing results of barcodes reveal a moderate decline in the composition of different barcodes during the expansion of iPSCs (colors are indicative for the corresponding fluorochrome of each barcode). The contribution of each specific barcode is represented as a percentage of the total number of barcode reads. Changes were analyzed at consecutive passages for each donor. ef In analogy, the composition of barcodes was tracked during differentiation towards and expansion of iMSCs (only successful for iPSCs of donors 2 and 3). Both donors revealed dominant subclones that emerged during generation of iMSCs. g Shannon Index measuring subclone-diversity. h CNVs in iPSCs and iMSCs of all three donors. SNP array analysis was performed with DNA samples of iPSCs (P1) and iMSCs (donor 1, P2; donor 2, P11; donor 3, P6). Chromosomal location, and the largest size of the variants (kbp) in either iPSCs or iMSCs are indicated for gains (black) and losses (gray) of more than 200 kbp. *Chromosomal locations where the corresponding mutation was present, but below the threshold, see also Additional file 6`

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