Reprogramming of Human Peripheral Blood Cells to Induced Pluripotent Stem Cells

Abstract

Direct reprogramming of human fibroblasts to induced pluripotent stem cells (iPS) has been achieved by ectopic expression of defined transcription factors. Derivation of human fibroblasts however is a time consuming process and requires punch biopsies or isolation of patient foreskin. Here we use a polycistronic vector encoding Oct4, Klf4, Sox2 and c-Myc to generate iPS cells from from frozen peripheral blood of several donors. Genomic DNA analyses indicated that iPS cells were derived from mature T cells as well as myeloid donor cells. Inducing pluripotency in peripheral blood would allow utilization of easy to get samples from the adult and, more importantly, provide convenient access to numerous patient samples stored in blood banks. The latter is of major interest as frozen blood samples, when reprogrammed to iPS cells, would allow the retrospective molecular analyses of rare diseases.

Generation of induced pluripotent stem cells using site-specific integration with phage integrase

“…Because the AF cells we used in this study could be mixtures of different cell populations, we could not identify the origin of the iPS cells by cell type. Recently, it has been reported that blood cells may be a good source for generating iPS cells (37)(38)(39)(40)(41).…”

Rejuvenating T cells

“…Several groups have generated T cell-derived iPS cells, [5][6][7][8] but not much is known about differentiating these cells back into antigen-specific T cells.…”

Promise and challenges of human iPSC-based hematologic disease modeling and treatment

“…Blood sample collection and storage are among the routine procedures in hospitals and clinics. Therefore, significant efforts have been made to develop efficient and safe procedures for reprogramming various types of blood cells (Table 1) [10,11,[13][14][15][16][17][18][19][20][21]. Umbilical cord blood (UCB) cell banks provide another rich source for cells with diverse HLA types.…”

“…T cells, which are abundant in peripheral blood and amenable to ex vivo expansion after stimulation of T cell receptor (TCR) activation, were the first subtypes of lineage-committed human blood cells being reprogrammed [16][17][18]. Analyses of the TCR genomic loci had shown the evidence of V(D)J recombination in resultant iPSCs, demonstrating their T cell origin [16][17][18]24]. These studies also provided the most convincing evidence that terminally differentiated human cells can be reprogrammed into a pluripotent state.…”