An improved protocol for generation and characterization of human-induced pluripotent stem cell-derived retinal pigment epithelium cells

• Published in: STAR protocols Vol. 3; no. 4; p. 101803
• Main Authors: Surendran, Harshini, Soundararajan, Lalitha, Reddy K, Vijay Bhaskar, Subramani, Janavi, Stoddard, Jonathan, Reynaga, Rene, Tschetter, Wayne, Ryals, Renee C., Pal, Rajarshi
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 16.12.2022; Elsevier
• Subjects: Cell culture; Cell Differentiation; Cell- and Tissue-Based Therapy; Health sciences; Humans; Induced Pluripotent Stem Cells; Macular Degeneration - therapy; Protocol; Retinal Pigment Epithelium; Stem cells; Cell culture; Health sciences; Cell differentiation; Stem cells
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2022.101803

We present an optimized protocol for guided differentiation of retinal pigment epithelium (RPE) cells from human-induced pluripotent stem cells (iPSC). De novo-generated RPE cells are mature, polarized, and mimic the cellular and molecular profile of primary RPE; they are also suitable for in vivo cell transplantation studies. The protocol includes an enrichment step, making it useful for large-scale GMP manufacturing. RPE cells produced following this protocol are appropriate for cell replacement therapy for macular degeneration and disease modeling. For complete details on the use and execution of this protocol, please refer to Surendran et al. (2021). [Display omitted] •The protocol is reliable, robust, and recapitulates in vivo retinal development•Enrichment step helps RPE production at a scale required for GMP grade manufacturing•RPE cells were authenticated by the expression of maturation markers•Subretinal transplantation of RPE resulted in significant vision rescue in RCS rats Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. We present an optimized protocol for guided differentiation of retinal pigment epithelium (RPE) cells from human-induced pluripotent stem cells (iPSC). De novo-generated RPE cells are mature, polarized, and mimic the cellular and molecular profile of primary RPE; they are also suitable for in vivo cell transplantation studies. The protocol includes an enrichment step, making it useful for large-scale GMP manufacturing. RPE cells produced following this protocol are appropriate for cell replacement therapy for macular degeneration and disease modeling.