The differentiation potential of precursor cells from the mouse lateral ganglionic eminence is restricted by in vitro expansion.

We have investigated whether the differentiation potential of attached cultures derived from the mouse lateral ganglionic eminence (LGE) is influenced by in vitro expansion. Primary neuronal cultures derived from the LGE give rise to neurons expressing the striatal projection neuron markers Islet1 (ISL1) and dopamine and cAMP-regulated phosphoprotein of 32 kilodaltons (DARPP-32) as well as the olfactory bulb interneuron marker Er81. Our previous results showed that after expansion in vitro, LGE precursor cells can be induced to differentiate into neurons which exhibit molecular characteristics of the LGE, such as the homeobox transcription factors DLX and MEIS2. We show here that while attached LGE cultures maintain Er81 expression through five passages, they lose the ability to generate ISL1- or dopamine and cAMP-regulated phosphoprotein of 32 kilodaltons-expressing neurons already after the first passage. This indicates that the expansion of LGE precursor cells restricts their differentiation potential in vitro. Interestingly, the undifferentiated LGE cultures retain the expression of both the Isl1 and Er81 genes, suggesting that precursor cells for both striatal projection neurons and olfactory bulb interneurons are present in these cultures. Thus the restriction in differentiation potential of the expanded LGE cultures likely reflects deficiencies in the differentiation conditions used

Regional specification of neurosphere cultures derived from subregions of the embryonic telencephalon.

We have studied the molecular specification of precursor cells in expanded neurosphere cultures derived from distinct subregions of the embryonic mouse telencephalon. These regionally derived cultures exhibited differential responses to the mitogens EGF and bFGF, suggesting that the precursors in these cultures were differentially specified as is the case in situ. To examine this further, cultures from each of the telencephalic subregions were expanded in both EGF and bFGF before differentiation. The neurons produced displayed molecular phenotypes similar to those normally derived from each of these regions in vivo. Moreover, analysis of gene expression in the undifferentiated cultures showed that the regionally derived neurospheres express many of the same developmental control genes as their in vivo counterparts. Taken together, the present findings suggest that precursor cells in neurosphere cultures, derived from distinct subregions of the embryonic telencephalon, maintain at least certain aspects of their molecular specification, even after significant expansion in vitro