In Vitro and in Vivo Enhanced Generation of Human A9 Dopamine Neurons from Neural Stem Cells by Bcl-XL.

Abstract

Human Neural Stem Cells derived from the ventral mesencephalon (VM) are powerful research tools and candidates for cell therapies in Parkinson′s disease (PD). Previous studies with VM dopaminergic neuron (DAn) precursors indicated poor growth potential and unstable phenotypical properties. Using the model cell line hVM1 (a new human fetal VM stem cell line) we have found that Bcl-XL enhances the generation of DAn from VM human neural stem cells. Mechanistically, Bcl-XL not only exerts the expected antiapoptotic effect, but it also induces proneural (NGN2, NEUROD1) and DA related transcription factors, resulting in a high yield of DAn with the correct phenotype of Substantia Nigra pars compacta (SNpc). The expression of key genes directly involved in VM/SNpc dopaminergic patterning, differentiation and maturation (EN1, LMX1B, PITX3, NURR1, VMAT2, GIRK2 and DAT) is thus enhanced by Bcl-XL. These effects on neurogenesis occur in parallel to a decrease in glia generation. These in vitro Bcl-XL effects are paralleled in vivo, after transplantation in hemiparkinsonian rats, where hVM1-Bcl-XL cells survive, integrate, and differentiate into mature DAn alleviating behavioral motor asymmetry. Bcl-XL then allows for human fetal VM stem cells to stably generate mature SNpc DAn both in vitro and in vivo, and is thus proposed as a helpful factor for the development of cell therapies for neurodegenerative conditions, PD in particular.