Expression of Neurodevelopmental Markers by Cultured Porcine Neural Precursor Cells

Abstract

Despite the increasing importance of the pig as a large animal model, little is known about porcine neural precursor cells. To evaluate the markers expressed by these cells, brains were dissected from 60-day fetuses, enzymatically dissociated, and grown in the presence of epidermal growth factor, basic fibroblast growth factor, and platelet-derived growth factor. Porcine neural precursors could be grown as suspended spheres or adherent monolayers, depending on culture conditions. Expanded populations were banked or harvested for analysis using reverse transcription-polymerase chain reaction (RT-PCR), immunocytochemistry, microarrays, and flow cytometry, and results compared with data from analogous human forebrain progenitor cells. Cultured porcine neural precursors widely expressed neural cell adhesion molecule (NCAM), polysialic acid (PSA)-NCAM, vimentin, Ki-67, and Sox2. Minority subpopulations of cells expressed doublecortin, β-III tubulin, synapsin I, glial fibrillary acidic protein (GFAP), and aquaporin 4 (AQP4) consistent with increased lineage restriction. A human microarray detected porcine transcripts for nogoA (RTN4) and stromal cellderived factor 1 (SDF1), possibly cyclin D2 and Pbx1, but not CD133, Ki-67, nestin, or nucleostemin. Subsequent RT-PCR showed pig forebrain precursors to be positive for cyclin D2, nucleostemin, nogoA, Pbx1, vimentin, and a faint band for SDF1, whereas no signal was detected for CD133, fatty acid binding protein 7 (FABP7), or Ki-67. Human forebrain progenitor cells were positive for all the genes mentioned. This study shows that porcine neural precursors share many characteristics with their human counterparts and, thus, may be useful in porcine cell transplantation studies potentially leading to the application of this strategy in the setting of nervous system disease and injury.

Development of cloned embryos from porcine neural stem cells and amniotic fluid-derived stem cells

“…In addition to nestin, a number of other markers have been associated with NS and progenitor cells. This study shows expression of NogoA, DCX, Cyclin D2, CD133, Hes1, Oct4, Desmin, CD-90, Nanog and Sox2 (Figure 2a-l), some of which have been reported as expressed by CNS precursor cells, or subsets thereof (Cai et al, 2002;Schwartz et al, 2003 and2005). The expression of these markers ( Figure 2a-l) by porcine NS cells argues for an important role for these genes in the neural development of large mammals.…”

“…NS cells (Gage, 2000;Jiang et al, 2002) can be cultured over the long term and are able to self-renew and differentiate into neurons, astrocytes and oligodendrocytes when exposed to cytokines in vitro Weiss, 1992 and1996;Schwartz et al, 2005). Thus, the NS cell is one type of well-defined somatic stem cell that can proliferate and differentiate in vitro.…”

AQP4 knockout impairs proliferation, migration and neuronal differentiation of adult neural stem cells

“…In the damaged brain, adult neurogenesis is also found in the neocortex, striatum, amygdala and substantia nigra (Gould, 2007) where AQP4 has a high level of expression. Finally, it is reported that AQP4 is the main subtype of aquaporin in adult neural stem cells (ANSCs) (Cavazzin et al, 2006;La Porta et al, 2006;Schwartz et al, 2005).…”

Progenitor Cells from the Porcine Neural Retina Express Photoreceptor Markers After Transplantation to the Subretinal Space of Allorecipients

“…In addition to anatomical considerations, we have recently reported that the gene expression profile of cultured porcine forebrain progenitor cells more closely resembles that of the analogous human cells than that of the mouse [16]. This raises the question of whether progenitor cells can also be cultured from the pig retina and, if so, whether the gene expression of these cells resembles their human counterparts [17,18] more than the analogous mouse cells [10].…”