Millipore Applications Bibliography |
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Lineage selection of functional and cryopreservable human embryonic stem cell-derived neurons. | ||
| Authors: | Ladewig, Julia, et al. | |
| Citation: | Stem Cells, 26: 1705-12 (2008) | |
| Pub Med ID: | 18420830 | |
| Year: | 2008 | |
| 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. | |
| Species: | Human | |
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