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Lineage selection of functional and cryopreservable human embryonic stem cell-derived neurons.

Author	Julia Ladewig, Philipp Koch, Elmar Endl, Banu Meiners, Thoralf Opitz, Sebastien Couillard-Despres, Ludwig Aigner, Oliver Brüstle
Citation Information	Stem cells (Dayton, Ohio), 26:1705-12 (2008)
Keywords	Animals, Cell Lineage, Cells, Cultured, Cryopreservation, Electrophysiology, Embryonic Stem Cells, Flow Cytometry, Green Fluorescent Proteins, Hippocampus, Humans, Mice, Microtubule-Associated Proteins, Neurons, Neuropeptides, Promoter Regions, Genetic
Species:	Human
Related Products	MAB377, MAB378
Pub Med ID	18420830

A major prerequisite for the biomedical application of human embryonic stem cells (hESC) is the derivation of defined and homogeneous somatic cell types. Here we present a human doublecortin (DCX) promoter-based lineage-selection strategy for the generation of purified hESC-derived immature neurons. After transfection of hESC-derived neural precursors with a DCX-enhanced green fluorescent protein construct, fluorescence-activated cell sorting enables the enrichment of immature human neurons at purities of up to 95%. Selected neurons undergo functional maturation and are able to establish synaptic connections. Considering that the applicability of purified hESC-derived neurons would largely benefit from an efficient cryopreservation technique, we set out to devise defined freezing conditions involving caspase inhibition, which yield post-thaw recovery rates of up to 83%. Combined with our lineage-selection procedure this cryopreservation technique enables the generation of human neurons in a ready-to-use format for a large variety of biomedical applications.