Differentiation of Porcine Inner Cell Mass Cells Into Proliferating Neural Cells

Abstract

Here we report the culture and differentiation of porcine neural cells derived from the inner cell masses (ICMs) of blastocyst-stage embryos. Manually dissected ICMs were cultured on feeder layers of inactivated mouse embryonic fibroblasts (MEFs). Neural rosette-like structures were selected and passaged mechanically. These structures were transient and disappeared after six passages. After transfer to Matrigel-coated dishes, the cells proliferated for approximately 2 months. Expression of neural progenitor cell (NPC)-specific genes NESTIN, SOX1, SOX2, and MUSASHI as detected by RT-PCR suggests that the cell culture contained neural progenitor-like cells. To further confirm the culture of neural progenitor-like cells, we analyzed their differentiation potential in vitro. The porcine neural cells were able to differentiate into glial fibrillary acidic protein (GFAP)-positive astrocytes and O4-positive oligodendrocytes, identified by quantitative RT-PCR and immunofluorescence. Differentiated cells expressed microtubule-associated protein 2 (MAP2) RNA but were unable to differentiate into neurons. It is concluded that porcine blastocysts have the ability to provide an expandable source of neural cells that can develop into glial subtypes.