Generation and Characterization of Aniridia iPSCs

Aniridia patient iPSCs were generated using non-integrating episomal reprogramming of dermal fibroblasts extracted from a skin biopsy from the patient’s arm, following established protocols.²⁸ (link)^,⁵³ (link) A minimum of 2 clonal lines were expanded and characterized as described previously.²⁹ (link)^,⁵³ (link) The control (WT) iPSCs used in this study have been published previously.⁵³ (link) The H9 ESC line was obtained from WiCell (hPSCreg WAe009-A). All iPSC lines were maintained in mTESR Plus medium (STEMCELL Technologies, Canada) with 0.1% penicillin/streptomycin (Pen/Strep) on Matrigel-coated wells (1:100) (Corning, USA). For passaging, ReLESR (STEMCELL Technologies) was used for detaching, and after 24 h, iPSCs were fed daily with mTESR Plus until confluent.

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Lima Cunha D., Sarkar H., Eintracht J., Harding P., Zhou J.H, & Moosajee M. (2023). Restoration of functional PAX6 in aniridia patient iPSC-derived ocular tissue models using repurposed nonsense suppression drugs. Molecular Therapy. Nucleic Acids, 33, 240-253.

Publication 2023

Pen/Strep Matrigel-coated wells ReLESR

Aniridia Biopsy Clonal Episomal Fibroblasts Ipscs Matrigel Patient Penicillin Skin Stemcell Streptomycin

Corresponding Organization :

Other organizations : University College London, Radboud University Nijmegen, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, The Francis Crick Institute, Moorfields Eye Hospital

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Variable analysis

independent variables

Aniridia patient iPSCs generated using non-integrating episomal reprogramming of dermal fibroblasts
Control (WT) iPSCs

dependent variables

Characteristics of iPSC clones (minimum of 2 clonal lines)

control variables

All iPSC lines maintained in mTESR Plus medium with 0.1% penicillin/streptomycin on Matrigel-coated wells
Passaging using ReLESR for detaching, and feeding daily with mTESR Plus until confluent

controls

The control (WT) iPSCs used in this study have been published previously
The H9 ESC line was obtained from WiCell (hPSCreg WAe009-A)

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