Establishing a centralized repository of human pluripotent stem cells for neurodegeneration research.

Abstract

In partnership with the National Institute on Aging (NIA), the National Centralized Repository for Alzheimer's Disease and Related Dementias (NCRAD) has been funded to establish a centralized repository of induced pluripotent stem cells (iPSCs). Investigators deposit iPSCs derived from patients with Alzheimer's disease and related dementias (ADRD), as well as iPSCs from healthy patients. However, much variability exists between iPSC lines, which is compounded by the diversity of culture conditions used by laboratories. Additionally, many labs do not have the resources to perform thorough quality control measures or to distribute high demand lines to other investigators. NCRAD strives to ease the distribution burden for investigators and provide standardized, quality controlled iPSCs to the ADRD community. Upon receipt, iPSCs are screened for multiple sources of contamination, including mycoplasma. Identity and APOE status are established utilizing a 94 single nucleotide polymorphism (SNP) genotyping assay. G-band karyotype is performed to assess genetic stability. Pluripotency and differentiation efficiency is analyzed by the hPSC Scorecard assay after embryoid body formation. Any pathogenic mutations are verified by Sanger sequencing or whole genome sequencing (WGS). WGS data will be shared and distributed by the National Institute on Aging Genetics of Alzheimer's Disease Storage Site (NIAGADS). Additionally, many lines contain additional phenotype data available at the National Alzheimer's Coordinating Center (NACC). To date, 156 iPSC lines have been deposited at NCRAD. All distributable lines were acclimated to a standardized culture condition of mTeSR™1 and Matirgel®. About 36% of lines represented sporadic ADRD while 27% were unrelated control lines. An additional 37% of lines contained known mutations, including matched isogenic mutants and controls. Mutations include PSEN1, PSEN2, APP, MAPT, and C9orf72. Out of 75 iPSC lines screened, the most common APOE genotype was APOE 3/3 (49.3%), followed by APOE 3/4 (22.6%). Through these efforts, NCRAD seeks to remove the burden of distributing iPSCs to ADRD researchers by collecting iPSCs from investigators worldwide, standardizing the culture conditions, and performing valuable quality control measures. It is NCRAD's goal to facilitate the study of neurodegenerative disorders through the distribution of uniform iPSCs to the ADRD community.