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Primary culture of chick, mouse or human neural crest cells.

Nature protocols (2011-10-01)
Heather Etchevers
ZUSAMMENFASSUNG

A highly enriched population of neural crest cells (NCCs) from amniote embryos, such as from chicks, mice and humans, is desirable for experiments in fate determination. NCCs are also useful for testing the functional effects of molecular changes underlying numerous human diseases of neural crest derivatives and for investigating their potential for therapeutic compensation. This protocol details embryonic microdissection followed by neural tube explantation. Conditions favoring NCC expansion and the maintenance of their stem cell-like properties are described. Although neural crest-like cells can be derived from a number of sites in the mature organism, full potential is best ensured by their purification from their source tissue at the outset of migration. Going from embryo to established cell line takes 4 d; the first is the most labor-intensive day, but minimal intervention is required thereafter.

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Sigma-Aldrich
Albumin aus Rinderserum, fatty acid free, low endotoxin, lyophilized powder, BioReagent, suitable for cell culture, ≥96% (agarose gel electrophoresis)
Sigma-Aldrich
LookOut® Mycoplasma PCR-Detektions-Kit, Optimized for use with JumpStart Taq DNA Polymerase, D9307.
Sigma-Aldrich
Hydrocortison, γ-irradiated, powder, BioXtra, suitable for cell culture
Sigma-Aldrich
apo-Transferrin human, γ-irradiated, powder, BioXtra, suitable for cell culture, ≥98%