Protocol for generating transgene-free naive human induced pluripotent stem cells from somatic cells using modified Sendai viral system

Sendai virus (SeV) vector represents a powerful tool for generating naive and primed human induced pluripotent stem cells (iPSCs) from somatic cells. Here, we present a protocol for the generation of transgene-free naive human iPSCs from human dermal fibroblasts (HDFs) and human peripheral mononuclear cells (PBMCs) using a modified SeV vector system. We describe steps for thawing the HDFs or PBMCs, reseeding HDFs, SeV vector infection, reseeding the SeV-infected HDFs or PBMCs on an irradiated mouse embryonic fibroblast (iMEF) plate, switching to t2iLGö+Y medium, passaging the generated naive iPSCs, removing the SeV vectors, and cryopreserving the naive iPSCs. For complete details on the use and execution of this protocol, please refer to Kunitomi et al.¹^,².