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Forschungsdatenbank PMU-SQQUID

Lineage selection of functional and cryopreservable human embryonic stem cell-derived neurons.
Ladewig, J; Koch, P; Endl, E; Meiners, B; Opitz, T; Couillard-Despres, S; Aigner, L; Brüstle, O;
Stem Cells. 2008; 26(7): 1705-1712.
Originalarbeiten (Zeitschrift)

PMU-Autor/inn/en

Aigner Ludwig
Couillard-Després Sébastien

Abstract

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.


Useful keywords (using NLM MeSH Indexing)

Animals

Cell Lineage

Cells, Cultured

Cryopreservation/methods*

Electrophysiology

Embryonic Stem Cells/cytology*

Flow Cytometry

Green Fluorescent Proteins/metabolism

Hippocampus/embryology

Humans

Mice

Microtubule-Associated Proteins/genetics*

Neurons/metabolism*

Neuropeptides/genetics*

Promoter Regions, Genetic


Find related publications in this database (Keywords)

human embryonic stem cells
lineage selection
human neurons
cryopreservation