Directed differentiation of human pluripotent cells to neural crest stem cells

Abstract

Multipotent neural crest stem cells (NCSCs) have the potential to generate a wide range of cell types including melanocytes; peripheral neurons; and smooth muscle, bone, cartilage and fat cells. This protocol describes in detail how to perform a highly efficient, lineage-specific differentiation of human pluripotent cells to a NCSC fate. The approach uses chemically defined media under feeder-free conditions, and it uses two small-molecule compounds to achieve efficient conversion of human pluripotent cells to NCSCs in ~15 d. After completion of this protocol, NCSCs can be used for numerous applications, including the generation of sufficient cell numbers to perform drug screens, for the development of cell therapeutics on an industrial scale and to provide a robust model for human disease. This protocol can be also be applied to patient-derived induced pluripotent stem cells and thus used to further the knowledge of human disease associated with neural crest development, for example, Treacher-Collins Syndrome.

Cell transplantation strategies for acquired and inherited disorders of peripheral myelin

“…As protocols for differentiating neural crest precursor cells from iPSCs are already established,28, 34, 35 we first examined iPSC‐derived human NCSCs as a candidate cell type for Schwann cell replacement therapy in demyelinated nerves. iPSCs from two normal subjects (lines 00i and 14i) were directly differentiated into human NCSCs via a protocol utilizing Wnt signaling activation and Activin/Nodal signaling inhibition (Fig.…”

<b>Induction of neural crest cells from human dental pulp-derived induced pluripotent stem </b><b>cells </b>

“…Further study is needed to determine whether larger neurosphere size would provide an advantage for NCC differentiation. We proceeded to examine the characteristics of the migrated cells from rosettes based on mRNA expression of NCC marker genes such as PAX3, P75, SLUG (SNAIL2), SNAIL (SNAIL1), and SOX9 (1,35,36,43,58). Expression levels of these genes in the migrated cells were significantly higher than those in original dental pulp-derived iPS cells.…”

Melanoma genotypes and phenotypes get personal

“…Neural crest-derived cells, by virtue of their relationship to an incredibly diverse assortment of human diseases, attract interest from many investigative perspectives beyond nevi and melanoma. 1 Indeed, it is now possible to direct cells into a neural crest stem cell fate to guide better understanding of diseases involving the craniofacial skeleton, cornea, teeth, and nervous systems. 1 Melanocytes are neural crest-derived and are responsible for producing melanin by virtue of uniquely possessing tyrosinase, the enzyme used for creating the distinguishing pigment.…”

“…1 Indeed, it is now possible to direct cells into a neural crest stem cell fate to guide better understanding of diseases involving the craniofacial skeleton, cornea, teeth, and nervous systems. 1 Melanocytes are neural crest-derived and are responsible for producing melanin by virtue of uniquely possessing tyrosinase, the enzyme used for creating the distinguishing pigment. The majority of nevi occurring after birth possess a mutation in the BRAF gene that encodes a serine/threonine kinase central to the mitogen-activated protein kinase (MAPK)-signaling pathway designated BRAFV600E.…”