Establishment of human iPSC line NCCSi003-A from CD34+cells of peripheral blood collected during apheresis of healthy donor from Indian ethnicity

We present generation of iPSCs from CD34+ cells isolated from peripheral blood, collected during apheresis of a healthy female individual. We nucleofected the CD34+cells by episomal vectors containing Oct4, Sox2, L-Myc, Lin28, Klf4 and p53DD (dominant negative mutation in p53). The resultant colonies showed cobble-stone appearance and stained positive for alkaline phosphatase. The colonies demonstrated presence of pluripotency markers by immunofluorescence, flow-cytometry and PCR. The plasmids were lost from cells subsequently during passages as assessed by PCR. Karyotype analysis demonstrated a stable genome. The cells had capability to differentiate to cells from all three-germ lineages in vitro

Development and characterization of human iPSC line NCCSi004-A from umbilical cord blood (UCB) derived CD34+cells obtained from donor belonging to Indian ethnic population

Here we report the reprogramming of CD34+ cells obtained from UCB of a healthy donor female child belonging to the Indian ethnic population. These CD34+cells were subjected to nucleofection for delivery of episomal vectors expressing Oct4, Sox2, L-Myc, Lin28, Klf4 and p53DD (negative mutation in p53). The iPSC colonies expressed pluripotency markers as detected by PCR, immunofluorescence and flow-cytometry. The removal of plasmid was confirmed by its absence in cells at higher passages. Karyotype analysis revealed a stable genome. The property of in vitro differentiation to tri-lineage was confirmed by expression of markers by immunofluorescence