PMC

(A) Embryoid bodies derived from PB34 and PBMC iPS cells yield hematopoetic colonies in semisolid methylcellulose media: burst forming unit-erythroid (BFU-E), colony forming unit-granulocyte (CFU-G), colony forming unit-macrophage (CFU-M), colony forming unit-granulocyte, macrophage (CFU-GM) and colony forming unit-granulocyte, erythroid, macrophage (CFU-GEMM). Total number of each type of colony was counted.
(B) Representative images of various types of hematopoietic colonies. Images were acquired with a standard microscope (Nikon, Japan) with a 20x objective.
(C, D) Hematoxylin and eosin staining of teratomas derived from immunodeficient mice injected with PB34 iPS (C) and PBMC iPS (D) cells show tissues representing all three embryonic germ layers.
(E) Genomic DNA from peripheral blood-derived iPS lines grown was digested with Nco I and analyzed for V(D)J rearrangements at the TCRδ (T-cell receptor Delta) locus by Southern blotting using a 3’Jδ3 probe.
(F) TCRδ V(D)J recombination of blood-derived iPS cell clones. Lanes 2–8 and lanes 13–17 are PBMC iPS lines. B-cell lines on lanes 8 and 9 showed no rearrangement. TCRδ rearrangement was observed for some PBMCs derived iPS lines (lanes 2–6 and 8). Lanes 1, 11, 12, 18, 19 are H1 hES cells, PB34 iPS cells, fibroblast cells, fibroblast derived iPS using retrovirus and lentivirus, respectively. The red arrow indicates expected size of the germline band. Orange arrow indicates re-arranged bands.
For further information on the pluripotency, V(D)J rearrangement and fingerprint analysis performed on the peripheral blood-derived iPSC clones see also .

(A) Scheme for reprogramming human peripheral blood (PB) mononuclear cells (PBMCs) and CD34+ cells (PB CD34+). Morphology of the typical peripheral blood cells and images of hES cell-like iPS colonies are shown.
(B) Images of PB34 iPS colonies. Bright field images were acquired with a standard microscope (Nikon, Japan) with a 10x objective. Immunohistochemistry of PB derived iPS cell colonies expressing markers for OCT4, NANOG, Tra-1-60 and alkaline phosphatase (AP). Hoechst staining indicates the total cell content per field. Fibroblasts surrounding human iPS colonies serve as internal negative controls for immunohistochemistry staining. Images were acquired with a standard microscope (Nikon, Japan) with a 10x objective.
(C) Images of PBMC (Donor GH) iPS colonies. Bright field images were acquired with a standard microscope (Nikon, Japan) with a 10x objective. Immunohistochemistry of PB derived iPS cell colonies expressing markers for OCT4, NANOG, Tra-1-60 and alkaline phosphatase (AP). Hoechst staining indicates the total cell content per field.
(D) Images of PBMC (Donor 34, 50, 76) iPS colonies. Bright field images were acquired with a standard microscope (Nikon, Japan) with a 10x objective. Immunohistochemistry of PB derived iPS cell colonies expressing marker for NANOG. DAPI staining indicates the total cell content per field.
(E) Quantitative reverse transcription–PCR analyses for the expression of ES cell-marker genes NANOG, hTERT, GDF3 and REX1 in PB CD34+ and PBMCs derived iPS cells and human H1 ES cells. Individual PCR reactions were normalized against β-ACTIN and plotted (Log₁₀ scale) relative to the expression level in the H1 ES cells, which was set to 1.
(F) Scatter plots comparing PB34 iPS and PBMC iPS cells global gene expression profiles to parental (Left) and H1 human ES cells (Right). The black lines indicate the linear equivalent and two-fold changes in gene expression levels between the paired cell types. Positions of pluripotency genes Oct4, Sox2, Nanog and Lin28 in scatter plots are indicated.
(G) Bisulfite genomic sequencing of the NANOG promoters reveals demethylation in the iPS cell lines. Each horizontal row of circles represents an individual sequencing reaction for a given amplicon. Open and filled circles represent unmethylated and methylated CpGs dinucleotides, respectively. Percentage of methylation is indicated for each cell line. For further characterization of the peripheral blood-derived iPSC clones see also .