Oral communication, PS3 / C38

Official XXIst International Pigment Cell Conference website - 21-24 Sept 2011, Bordeaux - France | updated: September 04 2011

Characterisation of pluripotent immortal postnatal mouse neural crest-like stem cells

We have established a novel type of pluripotent neural crest-like stem cells from neonatal mouse epidermis (Sviderskaya et al., 2009, FASEB J). These cells isolated as three independent immortal lines resemble neural crest stem cells in their apparent capacity to differentiate into several cell types normally derived from the neural crest. Using alternative regulatory factors, they could be converted to either melanoblasts and melanocytes, chondrocytes, Schwann precursor cells, or functional sensory neurons. Here we investigated whether neural crest-like stem cells can be induced to produce other cell types. Firstly, we examined whether neuronal cells can be produced in the conditions previously shown to induce production of pre-Schwann cells. We examined the effect of transforming growth factor β1 (TGFβ1) and neuregulin β1 (NRG1-β1) on these cells growing in RPMI medium with foetal calf serum (FCS). These factors significantly increased cell proliferation in 7 days with the highest effects resulting from treatment with both factors and NRG1-β1-treatment along (25.5% higher level of proliferation compared to TGFβ1-treatment) with increase of bi- and tri-polar cells. These cells showed immunoreactivity for a neuronal marker, neurofilament. About 14% of all cells showed positive staining with both growth factors after 7 days of treatment. The percentage of stained cells increased after longer treatments. Secondly, we confirmed that smooth muscle cells, another mesenchymal cell type and product of normal neural crest cells, could be produced by neural crest-like stem cells grown in the presence of BMP-2, TGFβ1 and FGF2 in DMEM medium with FCS. These factors increased cell proliferation with the effect of FGF2 being the highest and similar to that of all three factors combined. These factors also affected cell morphology. Immunostaining for phalloidin confirmed the appearance of triangular, stellate and spindle-shaped cells. They were immunoreactive for α-smooth muscle actin with the highest number of positive cells in BMP-2-treated cultures. Thus our immortal neural crest-like cells produce neuronal cells in the conditions previously described to induce production of Schwann precursor cells, and two types of mesenchymal cells, chondrocytes and smooth muscle cells, and thus can differentiate into six cell lineages. These lines provide a new and valuable model for large-scale studies of the biology and gene expression of neural crest.

Advertisement from our sponsor: