High-Fidelity Reprogrammed Human IPSCs Have a High Efficacy of DNA Repair and Resemble hESCs in Their MYC Transcriptional Signature

Figure 1

CB progenitors and CB-derived iPSCs closely resemble hESCs DNA repair gene expression signature. Microarray gene expression of selected DNA repair genes. (a)(i) Shown are hierarchical clustering heatmaps of mRNA from donor fibroblasts, donor CD34⁺ CB, and CB.iPSC derived with (+MSC) and without (−MSC) bone marrow stromal cell activation. hiPSC lines included sa-CBiPSC derived from stromal-activated CD34⁺ MP (; E5C3, E12C5, and E17C6: 6.2, 6.13, and 19.11), standard CB-iPSC, lines derived from CD34⁺ MP without stromal activation (, E17C1, E20C2, and E24C1), and standard CB-iPSC lines derived from CB unsorted mononuclear cells (, iCB9, iCB8, and iCB2.5). hESC lines included () H9, H7, and ES03. Signal intensities are from averaged independent biological replicate microarray samples ( as indicated). Expression array data depicts normalized values of the mean transcript levels for a subset of DDR genes in each group of the indicated cell lines. (a)(ii) Dot plots represent the normalized values of the signal intensities for PARP1/XRCC5/XRCC6 with corresponding values between categories indicated in the array data in (a)(i) ( as indicated in (a)(i)). Paired tests with significance () or without significance (NS; ) with values of control hESC are indicated (▲ = 4F CB.iPSC; ● = 7F CB.iPSC). (b)(i) Representative Western blot from the whole cell lysates of hESCs (H9 and ES03), CB (CD34⁺), two independent sa-CB-iPSCs (6.2 and E12C1), and adult fibroblasts (Ad.Fib) showing the steady state levels of PARP1 and Ku80 and ATM. β-Actin was used as the loading control. (b)(ii) Graphical representation of Western blots by ImageJ quantified-densitometry analysis normalized to β-actin () in hESC (H9, ES03, and H7), sa-CB-iPSC (CB6.2, CB6.13, and CB19.11), standard CB mononuclear CB-iPSC (iCB9, iCB8, and iCB2.5), CB (CD34⁺), and adult fibroblasts (Ad.Fib). Results are representative of the mean of two independent experiments of each set ± SEM, and , based on 2-way ANOVA (multiple comparisons test) on combined expression of genes.

(a)

(b)