Stem cells from farm animals are valuable cell models for the study of development, differentiation, and are potential efficient donors for nuclear transfer. Here we report the isolation and characterization of stem cells from porcine skin. These porcine skin-originated sphere (PSOS) cells expressed the neural progenitor marker, nestin, as well as genes that are critical for pluripotency such as Oct4 and Stat3. The PSOS cells proliferated actively in vitro and retained normal karyotype after long-term culture. When cultured in defined medium, they generated cells with characteristics of neurons and astrocytes. A subpopulation of cells differentiated into adipocyte-like cells when cultured in 10% fetal bovine serum. Clonal study demonstrated that PSOS exhibited clonal-generating capability. Clonal populations from individual stem cells could form neuron-, astrocyte-, and adipocyte-like cells upon inducted differentiation. Our findings represent the first report of skin-originated stem cells isolated from non-rodent animals. (C) 2004 Elsevier Inc. All rights reserved.