Differentiation and maturation of embryonal carcinoma-derived neurons in cell culture

Abstract

We have previously shown that retinoic acid-treated cultures of the P19 line of embryonal carcinoma cells differentiate into neurons, glia, and fibroblast-like cells (Jones-Villeneuve et al., 1982). We report here that the monoclonal antibody HNK-1 reacts with the neurons at a very early stage of their differentiation and is, therefore, an early marker of the neuronal lineage. Cells in differentiated P 19 cultures synthesized acetylcholine but not catecholamines, suggesting that at least some of the neurons are cholinergic. The neurons also carry high- affinity uptake sites for GABA but not for serotonin. In long-term cultures, neuronal processes differentiated into axons and dendrites, which formed synapses. This biological system should prove valuable for examining the development and maturation of cholinergic neurons, since their differentiation occurs in cell culture.

Blastema cells derived from New Zealand white rabbit’s pinna carry stemness properties as shown by differentiation into insulin producing, neural, and osteogenic lineages representing three embryonic germ layers

“…To induce neural differentiation, SLCs were treated with RA, which has been introduced as a neural inducer in mouse and human embryonal carcinoma cells (Andrews et al 1984;McBurney et al 1988;Staines et al 1994) and mouse ESCs in vitro (Rohweddel et al 1999;Pacherník et al 2005). Present results indicated that RA could induce neural differentiation in SLCs 7 days after treatment, as confirmed by detection of GT3 and GD3 neural markers.…”

Necdin Downregulates Cdc2 Expression to Attenuate Neuronal Apoptosis

“…Cytoplasmic and nuclear soluble fractions were prepared from CGNs as described previously (Taniguchi et al, 2000). Embryonal carcinoma P19 cells were induced to differentiate into neurons as described previously (McBurney et al, 1988). The equal amounts of proteins (25-30 g) were separated by 10% SDS-PAGE, blotted onto Immobilon membrane (Millipore, Billerica, MA), and incubated with antibodies against necdin (NC243, 1:1000), ␤-tubulin (1: 1000; MA Biomedicals, Irvine, CA), and phospho-specific Rb (Ser795, 1:300; New England Biolabs, Beverly, MA).…”

Cytoplasmic localization is required for the mammalian ELAV‐like protein HuD to induce neuronal differentiation

“…We ®rst tried to discover the physiological function of the neurone-speci®c Hu proteins using two neurogenic cell lines: rat pheochromocytoma cell line PC12, and mouse embryonal carcinoma cell line P19, which differentiate into neurone-like cells in response to nerve growth factor (NGF) and retinoic acid, respectively (Jones-Villeneuve et al 1982;Greene et al 1987;McBurney et al 1988). To examine the endogenous levels of Hu proteins in PC12 cells, we ®rst performed an immuno¯uorescence analysis using monoclonal antibody 16A11, which speci®cally recognizes the mammalian Hu proteins, except for HuR (Marusich et al 1994) (Fig.…”