Lineage selection of functional and cryopreservable human embryonic stem cell-derived neurons.
Stem Cells
; 26(7): 1705-12, 2008 Jul.
Article
en En
| MEDLINE
| ID: mdl-18420830
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.
Texto completo:
1
Banco de datos:
MEDLINE
Asunto principal:
Neuropéptidos
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Criopreservación
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Células Madre Embrionarias
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Proteínas Asociadas a Microtúbulos
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Neuronas
Límite:
Animals
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Humans
Idioma:
En
Año:
2008
Tipo del documento:
Article