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Expedited RPE‐derivation from human embryonic stem cells by partial PARP‐1 inhibition (LB55)
Author(s) -
Pennington Britney,
Clegg Dennis
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.lb55
Subject(s) - embryonic stem cell , nicotinamide , microbiology and biotechnology , poly adp ribose polymerase , stem cell , biology , cellular differentiation , cancer research , chemistry , polymerase , genetics , gene , biochemistry , enzyme
The death of the retinal pigmented epithelium (RPE) induces the leading cause of blindness in the elderly, a disease called age‐related macular degeneration (AMD). Human embryonic stem cells (hESCs) have the potential to generate a limitless source of RPE for cellular therapies, but spontaneous differentiation methods require several months to produce mature cells. In 2013, a novel directed‐differentiation protocol described efficient derivation of RPE in 14 days using nicotinamide as a key‐inducing agent in the chemical cocktail. This study investigates the hypothesis that nicotinamide expedites RPE‐derivation by inhibiting poly(ADP‐ribose) polymerase‐1 (PARP‐1) during the neuralization component of differentiation. 3‐aminobenzamide (3‐ABA), a non‐hydrolyzable nicotinamide analogue and PARP‐1 inhibitor, was tested in place of nicotinamide. Gene expression analysis by qPCR, morphological assessment, and pigment quantification demonstrate that 3‐ABA affects differentiation during the first 4 days of differentiation, ultimately yielding cells with similar, yet attenuated, levels of RPE‐marker gene expression compared to controls. This suggests that nicotinamide contributes to RPE differentiation from hESCs by partially inhibiting PARP‐1. Since other mechanisms are likely involved, future investigation aims to identify additional pathways contributing to this efficient differentiation of RPE cells, which will be important for generating cells for therapeutic applications. Grant Funding Source : Supported by Sigma Xi, Fight For Sight, and the California Institute for Regenerative Medicine