Human Pluripotent Stem Cell-derived Neural Crest Cells

Human Pluripotent Stem Cell derived Neural Crest Cells (Cat. #1670) from ScienCell Research Laboratories are differentiated from a human pluripotent stem cell line (H9), which is human blastocyst derived. The monolayer H9 efficiently generate Neural Crest Cells (NCC) using HPSC Neural Crest Medium (PSCNCM), a serum-free medium for rapid and efficient neural induction of human pluripotent stem cells (H9s). Synergistic inhibition of glycogen synthase kinase 3 (GSK3) and transforming growth factor β (TGF-β), and activation of BMP4 signaling differentiates H9 to homogenous neural crest cells (NCC) within 7 days.

The derived NCC are characterized by immunofluorescence with antibodies specific to Sox10 and p75. The cell population is highly pure: >70% of cells express Sox10. H9-NCC are cryopreserved at P1 and delivered frozen. Each vial contains >1 x 10⁶ cells in 1 ml volume. Cells are negative for mycoplasma, bacteria, yeast and fungi. After reviving, NCC can be maintained in Neural Crest Cell Medium (NCCM) as an adherent culture for a short time. NCC are multipotent and able to differentiate into various cell types such as peripheral neurons, smooth muscle cells, melanocytes and so on. Specific factors can be added after reviving to direct the cells to terminal differentiation. To differentiate the H9-NCC, medium containing specific growth factors and reagents should be used (not provided).

Recommended Medium

It is recommended to use the provided Neural Crest Cell Medium (NCCM) for plating H9-NCC and expanding them in the short term. Adding ROCK inhibitor Y-27632 in the first 12-18 hours after reviving improves cell viability and attachment in adherent cultures.