Differentiation of embryonic stem cell-derived dopaminergic neurons is enhanced by survival-promoting factors

Here, we describe the generation of viable and dopamin-producing neurons de rived from pluripotent mouse embryonic stem cells. Neurotrophic factors in combination with survival-promoting factors, such as interleukin-1 beta, gl ial cell line-derived neurotrophic factor, neurturin, transforming growth f actor-beta (3) and dibutyryl-cyclic AMP, significantly enhanced Nurr1 and t yrosine hydroxylase (TH) mRNA levels, whereas En-1, mash-1 and dopamine-2-r eceptor mRNA levels were not upregulated. In parallel, mRNA levels of the a nti-apoptotic gene bcl-2 were found to be upregulated at terminal stages. D ouble immunofluorescence analysis revealed increased numbers of TH- and dop amine transporter-, but not gamma -aminobutyric acid- and serotonin-positiv e neurons in relation to synaptophysin-labeled cells by survival-promoting factors. Moreover, high-performance liquid chromatography analysis showed d etectable levels of intracellular dopamine. We conclude that survival-promo ting factors enhance differentiation, survival and maintenance of dopaminer gic neurons derived from embryonic stem cells. (C) 2001 Elsevier Science Ir eland Ltd. All rights reserved.