Abstract
Human pluripotent stem cell (hPSC)-derived neural crest (NC) cells present a valuable tool for modeling aspects of human NC development, including cell fate specification, multipotency and cell migration. hpsc-derived Nc cells are also suitable for modeling human disease and as a renewable cell source for applications in regenerative medicine. Here we provide protocols for the step-wise differentiation of human embryonic stem cells (hEscs) or human induced pluripotent stem cells (hipscs) into neuroectodermal and Nc cells using either the Ms5 coculture system or a novel defined culture method based on pharmacological inhibition of bone morphogenetic protein and transforming growth factor-β signaling pathways. Furthermore, we present protocols for the purification and propagation of hPSc-Nc cells using flow cytometry and defined in vitro culture conditions. our protocol has been validated in multiple independent hESc and hipsc lines. The average time required for generating purified hPSc-Nc precursors using this protocol is 2-5 weeks.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 688-701 |
| Number of pages | 14 |
| Journal | Nature protocols |
| Volume | 5 |
| Issue number | 4 |
| DOIs | |
| State | Published - Apr 2010 |
| Externally published | Yes |
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)