title: Standardized generation and differentiation of neural precursor cells from human pluripotent stem cells.
abstract: Precise, robust and scalable directed differentiation of pluripotent stem cells is an important goal with respect to disease modeling or future therapies. Using the AggreWell™400 system we have standardized the differentiation of human embryonic and induced pluripotent stem cells to a neuronal fate using defined conditions. This allows reproducibility in replicate experiments and facilitates the direct comparison of cell lines. Since the starting point for EB formation is a single cell suspension, this protocol is suitable for standard and novel methods of pluripotent stem cell culture. Moreover, an intermediate population of neural precursor cells, which are routinely >95% NCAM(pos) and Tra-1-60(neg) by FACS analysis, may be expanded and frozen prior to differentiation allowing a convenient starting point for downstream experiments.
authors.affiliation: Laboratory of Molecular Biology, Division of Intramural Research, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: O A
authors.initials: OA
authors.lastname: Kozhich
authors.forename: R S
authors.initials: RS
authors.lastname: Hamilton
authors.forename: B S
authors.initials: BS
authors.lastname: Mallon
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods* / Q000592:standards*; D002454:Cell Differentiation; D005434:Flow Cytometry; D006801:Humans; D058953:Neural Stem Cells / Q000166:cytology* / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D012015:Reference Standards