Induced pluripotent stem cells (iPSC) were derived from a 42-year-old male with clinical features of type 1 diabetes (T1D). The donated tissue was infected with nonintegrating Sendai viruses containing the coding sequences of the following human genes: OCT4, SOX2, c-MYC, and KLF4. Cells express pluripotency markers and can differentiate in-vitro into cells of the three germ layers.
Human keratinocyte cells were plated at a density of 5X104 cells per well of a matrix-coated 24-well plate in ACF medium. Cells were transduced overnight with four pluripotency-associated factor-expressing Sendai viral vectors. Fresh ACF medium was placed over the cells for 3 days. Four days after vector infection, medium was changed to serum-free/feeder-free induced pluripotent stem cells (iPSC) medium (HEScGRO medium) supplemented with mTeSR-1 medium (25%, vol/vol). 3-4 weeks after vector infection, iPS-like clones were picked up and plated at a density of one colony per well in Matrigel-coated wells, of a 96-well plate, for further propagation.