Hair Follicle‐derived Neural Crest Stem Cells (HF‐NCSCs) as a Cell Source of Enteric Neurons

The enteric nervous system (ENS), which regulates most of the gastrointestinal functions, is derived from neural crest cells during early embryonic development. Abnormal migration and differentiation of neural crest cells may cause defective formation of the ENS, thus leading to enteric aganglionosis (absence of ganglia consisting of neurons and glial cells) in the distal colon. In the present study, we aimed to regenerate enteric neurons by transplanting hair follicle‐derived neural crest stem cells (HF‐NCSCs) to the aganglionic distal colon. We first isolated hair follicle bulges from the vibrissa whisker pad of mice, and cultured the bulges for 3 to 4 days. After HF‐NCSCs migrated onto the culture surface, the bulges were removed, and HF‐NCSCs were cultured in an expansion medium for another week. HF‐NCSCs were found to be able to differentiate into myocytes, neurons and glial cells in different differentiation media. Ten days after they were transplanted to the hindgut explants isolated from E13.5 mouse embryos, the transplanted HF‐NCSCs spread along the gut explant cultured ex vivo and differentiated into neurons. Immunostaining of the serial cross sections indicated most of the transplanted cells were located in the myenteric plexus between two muscle layers. HF‐NCSCs were also transplanted to the descending colon of an adult mouse in vivo after laparotomy. One week later, the cells spread along the colon and differentiate into Tuj1 and PGP9.5 immunoreactive cells. Ten days after transplantation in vivo, subtype inhibitory neurons immunoreactive to nNOS were found among the transplanted cells, and a small number of transplanted cells differentiated into excitatory neurons immunoreactive to ChAT. Additionally, almost all the transplanted cells were found in the myenteric plexus which is also the normal location of endogenous enteric neurons between the longitudinal muscle and circular muscle layers 6 days after transplantation. These results showed that (1) enough HF‐NCSCs were obtained for cell transplantation from a small number of skin hair follicles, and (2) HF‐NCSCs from hair follicles could serve as a cell source for stem cell‐based therapy. This work also gives insights into the feasibility of using autologous cell transplantation with a less invasive hair follicle isolation method on patients with ENS defects in the colon. Support or Funding Information The work was supported by a grant from the Research Grants Council of the Hong Kong Special Administrative Region, China (Project No.: 14175017). This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .