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Joe Phillips, Kyle A. Wallace, Sarah Dickerson, Michael Miller, Amelia D. Gerner, Jessica M. Martin, Lynda S. Wright, Elizabeth E. Capowski, Enio T. Perez, David M. Gamm; Retinal Laminae Formation and Synapse Development in Optic Vesicle-like Structures Isolated from Human Blood-derived iPS Cells. Invest. Ophthalmol. Vis. Sci. 2012;53(14):2692.
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© ARVO (1962-2015); The Authors (2016-present)
We sought to determine if human induced pluripotent stem cells (iPSCs) derived from peripheral blood could generate optic vesicle-like structures (OVs) with the capacity to stratify and express markers of intercellular communication.
Activated T-lymphocytes from a routine peripheral blood sample were reprogrammed by retroviral transduction to iPSCs. The T-lymphocyte-derived iPSCs (TiPSCs) were characterized for pluripotency and differentiated to OVs containing a highly enriched population of neuroretinal progenitor cells (NRPCs). TiPSC-OVs were then manually isolated, pooled, and cultured en masse to more mature stages of retinogenesis. Throughout the stepwise differentiation process, changes in anterior neural, retinal, and synaptic marker expression were monitored by PCR, immunocytochemistry, and/or flow cytometry.
TiPSCs generated OVs in abundance, which contained a near homogenous population of proliferating NRPCs. These NRPCs differentiated into multiple neuroretinal cell types, similar to OV cultures from human embryonic stem cells and fibroblast-derived iPSCs. In addition, portions of some TiPSC-OVs maintained a distinctive neuroepithelial appearance and spontaneously formed primitive laminae, reminiscent of the developing retina. Retinal progeny from TiPSC-OV cultures upregulated numerous genes and expressed proteins critical for synaptogenesis and gap junction formation, concomitant with the emergence of glia and the expression of thrombospondins in culture.
We demonstrate for the first time that human blood-derived iPSCs can generate retinal cell types, providing a highly convenient donor cell source for iPSC-based retinal studies. We also show that cultured human iPSC-OVs have the capacity to self-assemble into rudimentary neuroretinal structures and express markers indicative of chemical and electrical synapses.
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