Abstract
Fibroblasts can be collected from deceased individuals, grown in culture, reprogrammed into induced pluripotent stem cells (iPSCs), and then differentiated into a multitude of cell types, including neurons. Past studies have generated iPSCs from somatic cell biopsies from either animal or human subjects. Previously, fibroblasts have only been successfully cultured from postmortem human skin in two studies. Here we present data on fibroblast cell cultures generated from 146 scalp and/or 53 dura mater samples from 146 postmortem human brain donors. In our overall sample, the odds of successful dural culture was almost two-fold compared with scalp (OR = 1.95, 95% CI: [1.01, 3.9], p = 0.047). Using a paired design within subjects for whom both tissues were available for culture (n = 53), the odds of success for culture in dura was 16-fold as compared to scalp (OR = 16.0, 95% CI: [2.1–120.6], p = 0.0007). Unattended death, tissue donation source, longer postmortem interval (PMI), and higher body mass index (BMI) were associated with unsuccessful culture in scalp (all p<0.05), but not in dura. While scalp cells proliferated more and grew more rapidly than dura cells [F (1, 46) = 12.94, p<0.008], both tissues could be generated and maintained as fibroblast cell lines. Using a random sample of four cases, we found that both postmortem scalp and dura could be successfully reprogrammed into iPSC lines. Our study demonstrates that postmortem dura mater, and to a lesser extent, scalp, are viable sources of living fibroblasts for culture that can be used to generate iPSCs. These tissues may be accessible through existing brain tissue collections, which is critical for studying disorders such as neuropsychiatric diseases.
Highlights
Living fibroblasts can be cultured from human cadavers for use in induced pluripotent stem cell generation
Fibroblasts can be converted into induced pluripotent stem cells (iPSCs), which can further be differentiated into neurons [3,4]
Fibroblast Characterization and Proliferation When fibroblast cells were first generated from postmortem scalp and dura, we found that they initially began to differentiate more slowly than our lab’s previous experience with live skin-generated fibroblast cells, which differentiate in 5–7 days
Summary
Living fibroblasts can be cultured from human cadavers for use in induced pluripotent stem cell (iPSC) generation. This is valuable to test the biological fidelity of the mature cells derived from ordinarily inaccessible tissue and compared back to the primary cells from the same individual. A number of studies have generated cells with a neuronal phenotype from fibroblasts derived from living subjects, as previously reviewed [1]. Two groups to date have cultured fibroblasts from postmortem human dermal tissue samples [5], [6]. Meske and colleagues generated human fibroblast cultures from postmortem abdominal skin biopsies [5]. The validity of studies of fully differentiated neurons derived from iPSCs is based on the premise that the derived differentiated neurons exhibit most if not all of the biological characteristics of their natural counterparts
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