Mouse Epidermal Neural Crest Stem Cell (EPI-NCSC) Cultures

Summary

Here we show our method to isolate mouse epidermal neural crest stem cells (EPI-NCSC). Technique involves micro-dissecting whisker follicles, isolating the bulge and placeing it into tissue culture. EPI-NCSC start to emigrate from bulge explants onto the substratum within 3 - 4 days.

Abstract

EPI-NCSC are remnants of the embryonic neural crest in an adult location, the bulge of hair follicles. They are multipotent stem cells that have the physiological property to generate a wide array of differentiated cell types, including neurons, nerve supporting cells, smooth muscle cells, bone/cartilage cells and melanocytes. EPI-NCSC are easily accessible in the hairy skin and can be isolated as a highly pure population of stem cells. This video provides a detailed protocol for preparing mouse EPI-NCSC cultures from whisker follicles. The whisker pad of an adult mouse is removed, and whisker follicles dissected. The follicles are then cut longitudinally and subsequently transversely above and below the bulge region. The bulge is removed from the collagen capsule and placed in a culture plate. EPI-NCSC start to emigrate from the bulge explants 3 to 4 days later.

Protocol

Dissection of the Bulge from Adult Mouse Whisker Follicles

1. We use 10 weeks to 6 months old mice. Younger mice often yield more cells. Euthanize mouse and submerge entire animal into 1:1 mixture of betadine (iodine solution) and hydrogen peroxide (available from pharmacy) for about 3 minutes.
   
2. Squirt entire mouse, especially the facial region, with 75% ethanol, and carry mouse to dissection microscope in laminar flow hood.
3. Dissect whisker pads, carefully avoiding cutting into hair bulbs, and pool in HBSS.
4. Dissect whisker follicles and pool in HBSS at room temperature. Do this by holding skin next to hair follicle with a forcep....

Discussion

By virtue of their migratory ability, EPI-NCSC can be isolated as a highly pure population of stem cells, which can be expanded in vitro. As embryonic remnants in an adult location, EPI-NCSC are potentially attractive candidates for future cell replacement therapies, biomedical engineering and/or regenerative medicine. We have tested EPI-NCSC in a mouse model of spinal cord injury, where they show desirable traits. Through gene expression profiling by LongSAGE (www.ncbi.nlm.nih.gov/geo, series number GSE4680) we showed t.......