Abstract
Induced pluripotent stem cell (iPSC)-technology is an important platform in medicine and disease modeling. Physiological degeneration and disease onset are common occurrences in the aging population. iPSCs could offer regenerative medical options for age-related degeneration and disease in the elderly. However, reprogramming somatic cells from the elderly is inefficient when successful at all. Perhaps due to their low rates of replication in culture, traditional transduction and reprogramming approaches with centenarian fibroblasts met with little success. A simple and reproducible reprogramming process is reported here which enhances interactions of the cells with the viral vectors that leads to improved iPSC generation. The improved methods efficiently generates fully reprogrammed iPSC lines from 105–107 years old subjects in feeder-free conditions using an episomal, Sendai-Virus (SeV) reprogramming vector expressing four reprogramming factors.In conclusion, dermal fibroblasts from human subjects older than 100 years can be efficiently and reproducibly reprogrammed to fully pluripotent cells with minor modifications to the standard reprogramming procedures. Efficient generation of iPSCs from the elderly may provide a source of cells for the regeneration of tissues and organs with autologous cells as well as cellular models for the study of aging, longevity and age-related diseases.
Highlights
Aging is accompanied by a significant decline in physiologic functions in several organs, and by a dramatic increase in disabilities
All fibroblast lines were cultured in fibroblast medium, consisting of DMEM-GlutaMax supplemented with 10% heat-inactivated fetal bovine serum (FBS)(Gibco, Life Technologies), 1mM non-essential amino acids and 1% penicillin/streptomycin (Gibco, Life Technologies). Induced pluripotent stem cell (iPSC) cells were maintained in Essential 8 medium (A15171; Life Technologies) on vitronectin (A14700; Life Technologies) coated-plates
Young and centenarian fibroblasts were transduced with EmGFP Cytotune SeV vector (MOI = 3)
Summary
Aging is accompanied by a significant decline in physiologic functions in several organs, and by a dramatic increase in disabilities. A part of this decline is related to cell. A method for the efficient reprogramming of human centenarian somatic cells. Nestle SA provided support in the form of salaries for the authors JP and MK and did not provide any additional economical support for study design, data collection and analysis, decision to publish, or preparation of the manuscript. JP and MK contributions as they are articulated in the “author contribution"
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