Applying hydrodynamic pressure to efficiently generate induced pluripotent stem cells via reprogramming of centenarian skin fibroblasts

• Published in: PloS one 2019-04, Vol.14 (4), p.e0215490
• Main Authors: Vosough, Massoud, Ravaioli, Francesco, Zabulica, Mihaela, Capri, Miriam, Garagnani, Paolo, Franceschi, Claudio, Piccand, Julie, Kraus, Marine R-C, Kannisto, Kristina, Gramignoli, Roberto, Strom, Stephen C
• Format: Article
• Language: English
• Subjects: Adult; Age; Age Factors; Age related diseases; Aged, 80 and over; Aging; Artificial chromosomes; Autografts; Cell culture; Cell cycle; Cells, Cultured; Cellular Reprogramming; Cellular Reprogramming Techniques - methods; Centenarians; Degeneration; DNA damage; EDTA; Efficiency; Fibroblasts; Fibroblasts - physiology; Genetic Vectors - genetics; Genomes; Geriatrics; Health aspects; Health sciences; Humans; Hydrodynamic pressure; Hydrodynamics; Induced Pluripotent Stem Cells - physiology; Infant, Newborn; Laboratories; Medicin och hälsovetenskap; Medicine; Methods; Nuclear reprogramming; Older people; Organs; Pathology; Physiological aspects; Pluripotency; Primary Cell Culture; Regeneration; Regenerative medicine; Reproducibility of Results; Sendai virus - genetics; Senescence; Skin; Skin - cytology; Skin Aging - physiology; Somatic cells; Stem cells; Technology; Transfection - methods; Transplantation, Autologous - methods; Viruses
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0215490

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.