3D-aggregated dermal stem cells with partial-pluripotency

Current stem cell therapy is expected to be an efficient method for replacement of lost cells in many intractable diseases, and several types of cells are considered candidate sources including iPSCs and ESCs. However, these cells still need to elucidate several serious problems. We are trying to establish safe pluripotent stem cells for early use to clinic. Here we have identified and established 3D spheroidal dermal stem cells, which possess partial pluripotency. We cultured the dermal cells from GFP carrying C57BL/6 male mice. After their second passage, we dissociate and quickly aggregate these cells and cultured on U-bottom 96-well low-cell-adhesion plates. The cells self aggregated in 3D structure. We examined pluripotent markers. We found that 3D-aggregated sphere expressed Nanog, Oct4, Sox2 and SSEA-1 protein. We named these 3D sphere as spheroidal dermal stem cells (sDSCs). Then we tried to differentiate sDSCs to ectodermal-lineage cells on PA6 cells. We found the appearance of neuron-like cells, having the neuronal markers such as Neurofilament and S-100b positive. For endodermal-lineage cell induction, sDSCs were cultured on collagen-coated dishes with FBS and Activin A. The differentiated cells express α-fetoprotein and Pdx-1. This study shows that sDSCs would be the low risk and potential applications of cell transplantation therapy and be able to breakthrough in stem cell treatment.