A longevity-specific bank of induced pluripotent stem cells from centenarians and their offspring

Abstract

Centenarians provide a unique lens through which to study longevity, healthy aging, and resiliency. Moreover, models of human aging and resilience to disease that allow for the testing of potential interventions are virtually non-existent. We obtained and characterized over 96 centenarian and offspring peripheral blood samples including those connected to functional independence data highlighting resistance to disability and cognitive impairment. Targeted methylation arrays were used in molecular aging clocks to compare and contrast differences between biological and chronological age in these specialized subjects. Isolated peripheral blood mononuclear cells (PBMCs) from 20 of these subjects were then successfully reprogrammed into high-quality induced pluripotent stem cell (iPSC) lines which were functionally characterized for pluripotency, genomic stability, and the ability to undergo directed differentiation. The result of this work is a one-of-a-kind resource for studies of human longevity and resilience that can fuel the discovery and validation of novel therapeutics for aging-related disease.

Keywords: centenarians; induced pluripotent stem cells; longevity.

FIGURE 1

Comparisons of biological versus chronological age in EL subjects. Parallel age estimates from five well‐established models of epigenetic aging for EL PBMCs. The colored lines represent the best linear fit from each model, while the dotted black lines show a theoretical perfect linear correlation for reference, where applicable. Each data point represents an individual within our bank. The top three panels show models trained on chronological age including the original 2013 pan‐tissue Horvath (a), Hannum (b), and ENCen40+ (c) clocks. (d and e) show predictions from models (PhenoAge and DunedinPACE) trained with the additional aid of clinical and phenotypic measurements. These clocks return more novel measurements that align more closely to a putative biological age, or rate of age acceleration, as opposed to chronological age.

FIGURE 2

Representative characterization of EL‐specific iPSCs. (a) Representative images of iPSC lines under brightfield (left), DAPI (middle), and TRA‐1‐81 (right). Images taken at 10× magnification. (b) Table of iPSC lines generated including demographic information (age, sex) and karyotype outcome based on g‐band analyses. (c) Representative karyotype reports of male EL iPSCs showing a normal karyotype (left) and mosaic loss of y chromosome (right). (d) Representative images of teratoma mass hematoxylin and eosin stains from EL iPSC lines representing mesoderm (arrows), endoderm (asterisks), and ectoderm (accent) tissue. Scale bars: 100 μM.

FIGURE 3

Forward programming of EL‐specific iPSCs into cortical neurons. Representative images from EL iPSC lines brought through forward programming to forebrain cortical neurons. Maturation markers TUJ1 (green), MAP2 (red), NUEN (cyan), and DAPI (blue) are shown. Scale bars: 100 μM.