Open AccessLineage Selection of Functional and Cryopreservable Human Embryonic Stem Cell‐Derived Neurons
Author(s) -
Ladewig Julia,
Koch Philipp,
Endl Elmar,
Meiners Banu,
Opitz Thoralf,
CouillardDespres Sebastien,
Aigner Ludwig,
Brüstle Oliver
Publication year - 2008
Publication title -
stem cells
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.159
H-Index - 229
eISSN - 1549-4918
pISSN - 1066-5099
DOI - 10.1634/stemcells.2008-0007
Subject(s) - biology , embryonic stem cell , cell sorting , microbiology and biotechnology , lineage (genetic) , stem cell , neurogenesis , negative selection , somatic cell , transfection , computational biology , cell , cell culture , genetics , gene , genome
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. Disclosure of potential conflicts of interest is found at the end of this article.