Standardized Generation and Differentiation of Neural Precursor Cells from Human Pluripotent Stem Cells
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- Kozhich, O.A., Hamilton, R.S. & Mallon, B.S. Stem Cell Rev and Rep (2013) 9: 531. doi:10.1007/s12015-012-9357-8
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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% NCAMpos and Tra-1-60neg by FACS analysis, may be expanded and frozen prior to differentiation allowing a convenient starting point for downstream experiments.