I provide detailed protocols for conduction and troubleshooting the key steps in our three most used experimental designs: (1) prospectively counting and sorting of human neural stem cells (NSCs)/committed progenitors before placing them in culture; (2) high-throughput methods of quantifying changes in NSC/progenitor proliferation, in vitro; and (3) retrovirally tagging NSCs before differentiation to assess cell fates in individual clones. Detailed troubleshooting of immunohistochemical and fluorescence-activated cell sorting staining is described. Some of these techniques overlap with other chapters in this volume. Ultimately, this provision of complementary technical information should help ensure the reader’s experimental success.
Key WordsHuman retrovirus packaging cell line neural stem cell immunochemistry BrdU fluorescence-activated cell sorting proliferation antibody differentiation ELISA