Kit-based remote collection and isolation of human reprogrammable somatic cells for generation of induced pluripotent stem cells and cerebral organoids.

Developing reproducible human disease models is a central goal of biomedical research and personalized medicine. Induced pluripotent stem (iPS) cell-derived models enable investigation of underlying cellular and molecular mechanisms in humans. However, obtaining reprogrammable somatic cells is a challenge, especially when patients have limited access to hospital and research facilities. Here we provide a step-by-step protocol for noninvasive isolation of human keratinocytes via trypsinization from scalp hair with intact follicles. The samples can be collected remotely by the donor, a family member or a healthcare provider using a kit and shipped long distance at ambient temperature. This offers a major advantage over widely used methods for iPS cell generation from other somatic cells, for example, dermal fibroblasts or peripheral blood mononuclear cells (PBMCs) (from invasive skin biopsies or blood collection), or renal epithelial cells (from urine). Keratinocytes can be reprogrammed to iPS cells that give rise to all three cell lineages, including the neural lineage, and used to generate cerebral organoids. The protocol yields iPS cells within 2 months of receiving the hair samples and requires basic familiarity with mammalian cell culture techniques without the need for specialized equipment. Generation of cerebral organoids from iPS cells requires 30-40 d and specialized equipment (an orbital shaker) for culture.