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Kyle Wallace, Amelia D. Verhoeven, E. Ferda Percin, Sara Dickerson, Michael Miller, Joe Phillips, Wei Shen, Elizabeth E. Capowski, Lynda S. Wright, David M. Gamm; Targeted Retinal Differentiation of Human iPS Cells Derived from Peripheral Blood Samples. Invest. Ophthalmol. Vis. Sci. 2011;52(14):2216.
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Human induced pluripotent stem cells (hiPSCs) can be produced from many somatic cell types, but perhaps the most noninvasive and convenient cell source is peripheral blood lymphocytes. However, the capacity for blood-derived hiPSCs to yield retinal progeny has not been evaluated. We sought to reprogram donor T-lymphocytes to hiPSCs and determine their potential to differentiate along the retinal lineage.
T-lymphocytes were isolated and expanded from a 3 mL sample of human whole peripheral blood (1). Reprogramming was performed using MMLV and the following transgenes: SOX2, OCT4, c-MYC, KLF4, NANOG and LIN28. The resulting hiPSC lines were characterized to assure pluripotency, whereupon selected lines were subjected to an established retinal differentiation protocol (2). Differentiating blood-derived hiPSC lines were then examined for the expression of early anterior neuroepithelial, eye field, retinal progenitor, RPE, and photoreceptor markers using RT-PCR, quantitative PCR, and ICC.
Multiple hiPSC lines were derived from peripheral blood samples that expressed appropriate pluripotency markers. Upon differentiation, these hiPSCs generated primitive anterior neuroepithelial/eye field phenotypes expressing such markers as PAX6, OTX2, SIX6 and LHX2 within the first 10 d. Further maturation led to the co-expression of the neural retinal progenitor marker CHX10 and Ki67 in a subset of neurospheres, which subsequently differentiated into more mature retinal cell types. RPE-like cells expressing MITF and ZO-1 also appeared within blood-derived hiPSC cultures by day 40.
The competence of blood-derived hiPSCs to produce retinal cell types was established in this study. The ability to generate and enrich for retinal populations and RPE from reprogrammed T-lymphocytes offers a simple, relatively noninvasive means of obtaining cells of potential use in the study of retinal disease.1. Brown (2010) PLoS ONE 5(6):e11373.2. Meyer (2009) PNAS 106:16698.
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