Figure 2. Generation of neural stem cells from adult peripheral CD34+ cells.

Adult peripheral CD34+ cells were transduced using Sendai virus constructs containing Oct3/4, Sox2, Klf4 and c-Myc. (A) A few adherent cells were observed in uninfected CD34+ cell cultures (Ctrl) in neural stem cell medium. These cells did not show any proliferation and or nestin immunostaining. (B) Following transduction and culture in neural stem cell medium, CD34+ cells resulted in more adherent proliferating cells, which were SOX-2 (red), nestin (green) and PAX6 (green) positive, but negative for OCT4 (red). Cells were counterstained with DAPI (blue). Gene expression profiling were studied using samples from one CD34+ cells culture (two technical repeats), one iPS differentiated neural stem cells culture (iPS-iNS, two technical repeats), two induced neural stem cells cultures (iNS, one with two technical repeats) and three human primary fetal neural progenitor cells cultures (NPC, one with two technical repeats). Sample-to-sample relationships based on covariance-based Principal Component Analysis (i) and correlation-based clustering analysis (ii) using 311 markers for pluripotency (C) and 197 neuronal progenitor markers (D) (NCBI PMID: 23117585).  Both analyses were performed in R (http://cran.r-project.org/) using the princomp and heatmap.2 functions respectively.  Samples depicted in the PCA scatterplot are represented as circles and described by type using color (CD34 = blue, iPSC-iNS = pink, iNS = green, NPC = red).  Marker expression is of type RMA (log, base=2).