NIH Research Festival
Background: Induced pluripotent stem cells (iPSC) have great potential in treating a variety of diseases, for instance, therapy using iPSC-derived Retinal Pigment Epithelial (RPE) cells to treat Age-related Macular Degeneration (AMD) displayed promising results. National Eye Institute (NEI) initialized a phase I clinical trial to treat dry AMD using iPSC-derived RPE cells on a scaffold patch, and Center for Cellular Engineering (CCE) of Clinical Center, NIH developed standard operating procedure (SOP) for manufacturing GMP grade iPSCs to support the clinical trial of NEI. Method: A clinical protocol recruiting AMD patients or age-matched healthy donors was initiated by investigators in NEI. Donors or AMD patients were screened and eligibility was determined in DTM of Clinical Center. Peripheral blood was collected and peripheral blood mononuclear cells (PBMCs) were isolated and cryopreserved. After thawing, CD34+ cells were selected and expanded; the expanded cells were reprogramed into iPSC using reprogramming cocktail of multiple plasmids (which plasmids??) that were delivered to the cells using a Nucleofector device. During reprogramming, cells that conformed to the specifications on iPSC were picked and expanded until the passage 10 cell stock was established. Results and Discussion:A robust procedure of manufacturing GMP grade iPSC under feeder-free and chemically defined Xeno-free conditions, consisting of detailed bioprocessing, and quality control (QC) measures, was established in CCE. The Critical Quality Attributes (CQA) and Critical Processing Parameters (CPP) for the generation of iPSC are outlined here. Using this method, we were able to consistently generate clinical-grade iPSC that will be used in a NEI-sponsored clinical trial to treat age-related macular degeneration. The methods outlined here may have a broader application for others that will require iPSC starting material.
Scientific Focus Area: Stem Cell Biology
This page was last updated on Friday, March 26, 2021