title: Induction of pluripotent stem cells from autopsy donor-derived somatic cells.
abstract: Human induced pluripotent stem cells (iPSCs) have become an intriguing approach for neurological disease modeling, because neural lineage-specific cell types that retain the donors' complex genetics can be established in vitro. The statistical power of these iPSC-based models, however, is dependent on accurate diagnoses of the somatic cell donors; unfortunately, many neurodegenerative diseases are commonly misdiagnosed in live human subjects. Postmortem histopathological examination of a donor's brain, combined with premortem clinical criteria, is often the most robust approach to correctly classify an individual as a disease-specific case or unaffected control. In this study, we describe iPSCs generated from a skin biopsy collected postmortem during the rapid autopsy of a 75-year-old male, whole body donor, defined as an unaffected neurological control by both clinical and histopathological criteria. These iPSCs were established in a feeder-free system by lentiviral transduction of the Yamanaka factors, Oct3/4, Sox2, Klf4, and c-Myc. Selected iPSC clones expressed both nuclear and surface antigens recognized as pluripotency markers of human embryonic stem cells (hESCs) and were able to differentiate in vitro into neurons and glia. Statistical analysis also demonstrated that fibroblast proliferation was significantly affected by biopsy site, but not donor age (within an elderly cohort). These results provide evidence that autopsy donor-derived fibroblasts can be successfully reprogrammed into iPSCs, and may provide an advantageous approach for generating iPSC-based neurological disease models.
authors.affiliation: Neurogenomics Division, The Translational Genomics Research Institute (TGen), Phoenix, AZ, USA.
authors.forename: Brooke E
authors.initials: BE
authors.lastname: Hjelm
authors.forename: Jon B
authors.initials: JB
authors.lastname: Rosenberg
authors.forename: Szabolcs
authors.initials: S
authors.lastname: Szelinger
authors.forename: Lucia I
authors.initials: LI
authors.lastname: Sue
authors.forename: Thomas G
authors.initials: TG
authors.lastname: Beach
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Huentelman
authors.forename: David W
authors.initials: DW
authors.lastname: Craig
mesh_terms: D000368:Aged; D001344:Autopsy; D018929:Cell Culture Techniques / Q000379:methods*; D054337:Cell Dedifferentiation / Q000235:genetics; D019070:Cell Lineage / Q000235:genetics; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000502:physiology*; D000090062:Kruppel-Like Factor 4; D008297:Male; D061251:Primary Cell Culture; D014019:Tissue Donors; D014161:Transduction, Genetic / Q000379:methods*