Patent ID: 6830927
Filing Date: 2004-12-14
Classification: C12N

Abstract:
A method for generating mammalian neural crest stem cells comprising:(a) obtaining mammalian, p75 immunonegative neuroepithelial stem cells derived from the neural tube from a mammalian embryo at a stage of embryonic development after closure of the neural tube by: (i) removing a sample of neural tube tissue from a mammal at a stage of embryonic development after closure of the neural tube; (ii) dissociating cells comprising the sample of neural tube tissue removed from the mammal; (iii) plating the dissociated cells in feeder-cell-independent culture on a substratum and in a media comprising fibroblast growth factor and chick embryo extract; and (iv) identifying p75 immunonegative cells via immunohistochemistry and obtaining the mammalian, p75 immunonegative neuroepithelial stem cells; (b) harvesting the mammalian, p75 immunonegative neuroepithelial stem cells by trypsinization; and (c) replating the harvested mammalian, p75 immunonegative neuroepithelial stem cells onto a fibronectin substrate and in a media comprising chick embryo extract, NGF, FGF and EGF to generate p75 immunoreactive, neural crest stem cells.