Neural crest stem cells from human epidermis of aged donors maintain their multipotency in vitro and in vivo

Abstract

Neural crest (NC) cells are multipotent stem cells that arise from the embryonic ectoderm, delaminate from the neural tube in early vertebrate development and migrate throughout the developing embryo, where they differentiate into various cell lineages. Here we show that multipotent and functional NC cells can be derived by induction with a growth factor cocktail containing FGF2 and IGF1 from cultures of human inter-follicular keratinocytes (KC) isolated from elderly donors. Adult NC cells exhibited longer doubling times as compared to neonatal NC cells, but showed limited signs of cellular senescence despite the advanced age of the donors and exhibited significantly younger epigenetic age as compared to KC. They also maintained their multipotency, as evidenced by their ability to differentiate into all NC-specific lineages including neurons, Schwann cells, melanocytes, and smooth muscle cells (SMC). Notably, upon implantation into chick embryos, adult NC cells behaved similar to their embryonic counterparts, migrated along stereotypical pathways and contributed to multiple NC derivatives in ovo . These results suggest that KC-derived NC cells may provide an easily accessible, autologous source of stem cells that can be used for treatment of neurodegenerative diseases or as a model system for studying disease pathophysiology and drug development.

On the road again: Establishment and maintenance of stemness in the neural crest from embryo to adulthood

“…Human NC stem cells have also been derived from direct reprogramming of fibroblasts by introduction of transcription factors, such as Sox10 230 or FoxD3, 231 which both play roles in maintenance of stem cell properties in NC cells (section 3). More recently NC cells were derived from keratinocytes of the interfollicular epidermis (another easily accessible source) of neonatal and adult donors by using growth factors for reprogramming 232,233 . These derived human NC stem cells have a molecular profile compatible with NC cell identity in animal models, they express genes such as Sox10, FoxD3, and AP2α and give rise to several NC derivatives such as neurons, glia, melanocytes, cartilage, and bone 46,225‐233 .…”

“…More recently NC cells were derived from keratinocytes of the interfollicular epidermis (another easily accessible source) of neonatal and adult donors by using growth factors for reprogramming 232,233 . These derived human NC stem cells have a molecular profile compatible with NC cell identity in animal models, they express genes such as Sox10, FoxD3, and AP2α and give rise to several NC derivatives such as neurons, glia, melanocytes, cartilage, and bone 46,225‐233 . Although future work will need to determine how similar the NC cells formed by these various methods are to each other and how accurately they represent endogenous NC development, it is clear that these approaches hold great promise for regenerative therapy purposes.…”

Neural crest-like stem cells for tissue regeneration

“…Therefore, this approach provides a rapid method of obtaining NCLSCs that can be potentially administered clinically. To date, NCLSCs have been derived from human and mouse fibroblasts, 177,186‐188 keratinocytes, 189,190 and melanocytes 191 via the introduction of transcription factors, such as SOX10 and FOXD3 , specific growth factors, or forced expression of Notch1 signaling, respectively. It has been reported that the derived NCLSCs are functional in vivo when they were investigated for neural repair in a zebrafish model 186 and migration capability in a chick embryo model system 177,191 ; however, more detailed and comprehensive analysis of in vivo functional outcomes is still necessary.…”

hReg-CNCC reconstructs a regulatory network in human cranial neural crest cells and annotates variants in a developmental context

“…For example, chr7:134379901-134380671 was predicted to regulate CALD1 . CALD1 was known to play an essential role in the regulation of smooth muscle and non-muscle contraction 59 , which was a derivative of neural crest 60 . chr17:597438-597652 regulated FAM57A , which was linked to “Sclerosteosis 1”, a disease with abnormal character of skull and mandible 61 .…”