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

Stem Cells
Julia LadewigO Brüstle

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.

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Citations

Nov 19, 2013·Nature Neuroscience·Julia LadewigOliver Brüstle
May 25, 2011·Proceedings of the National Academy of Sciences of the United States of America·Sangmi ChungKwang-Soo Kim
Nov 23, 2011·Proceedings of the National Academy of Sciences of the United States of America·Jason P WeickSu-Chun Zhang
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Jan 5, 2011·BMC Neuroscience·Jingzhong ZhangSebastien Couillard-Despres
Feb 27, 2010·Stem Cell Reviews and Reports·Stefanie TersteggeOliver Brüstle
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Mar 15, 2011·The European Journal of Neuroscience·Sebastien Couillard-Despres, Ludwig Aigner
Aug 19, 2015·Molecular and Cellular Neurosciences·Brooke A DeRosaDerek M Dykxhoorn
May 23, 2013·Stem Cells Translational Medicine·Masato MaruyamaTetsuo Sugimoto
Mar 3, 2019·International Journal of Molecular Sciences·Sarah SchwarzAnnika Lehmbecker

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