Article

Stem Cell Reviews and Reports

, Volume 7, Issue 4, pp 799-814

Open Access This content is freely available online to anyone, anywhere at any time.

Human Epidermal Neural Crest Stem Cells (hEPI-NCSC)—Characterization and Directed Differentiation into Osteocytes and Melanocytes

  • Oliver ClewesAffiliated withInstitute of Genetic Medicine, and North East England Stem Cell Institute (NESCI), Centre for Life, Newcastle University
  • , Alla NarytnykAffiliated withInstitute of Genetic Medicine, and North East England Stem Cell Institute (NESCI), Centre for Life, Newcastle University
  • , Kevin R. GillinderAffiliated withInstitute of Genetic Medicine, and North East England Stem Cell Institute (NESCI), Centre for Life, Newcastle University
  • , Andrew D. LoughneyAffiliated withMaternity Unit, Women’s Services, Newcastle upon Tyne Hospitals NHS Foundation Trust, Royal Victoria Infirmary
  • , Alison P. MurdochAffiliated withNewcastle Fertility Centre at Life, BioScience Centre, International Centre for Life, Times Square
  • , Maya Sieber-BlumAffiliated withInstitute of Genetic Medicine, and North East England Stem Cell Institute (NESCI), Centre for Life, Newcastle UniversityInstitute of Genetic Medicine Centre for Life, Newcastle University Email author 

Abstract

Here we describe the isolation, characterisation and ex-vivo expansion of human epidermal neural crest stem cells (hEPI-NCSC) and we provide protocols for their directed differentiation into osteocytes and melanocytes. hEPI-NCSC are neural crest-derived multipotent stem cells that persist into adulthood in the bulge of hair follicles. Multipotency and self-renewal were determined by in vitro clonal analyses. hEPI-NCSC generate all major neural crest derivatives, including bone/cartilage cells, neurons, Schwann cells, myofibroblasts and melanocytes. Furthermore, hEPI-NCSC express additional neural crest stem cell markers and global stem cell genes. To variable degrees and in a donor-dependent manner, hEPI-NCSC express the six essential pluripotency genes C-MYC, KLF4, SOX2, LIN28, OCT-4/POU5F1 and NANOG. hEPI-NCSC can be expanded ex vivo into millions of stem cells that remain mulitpotent and continue to express stem cell genes. The novelty of hEPI-NCSC lies in the combination of their highly desirable traits. hEPI-NCSC are embryonic remnants in a postnatal location, the bulge of hair follicles. Therefore they are readily accessible in the hairy skin by minimal invasive procedure. hEPI-NCSC are multipotent somatic stem cells that can be isolated reproducibly and with high yield. By taking advantage of their migratory ability, hEPI-NCSC can be isolated as a highly pure population of stem cells. hEPI-NCSC can undergo robust ex vivo expansion and directed differentiation. As somatic stem cells, hEPI-NCSC are conducive to autologous transplantation, which avoids graft rejection. Together, these traits make hEPI-NCSC novel and attractive candidates for future cell-based therapies and regenerative medicine.

Keywords

Neural crest Adult stem cell Human epidermal neural crest stem cell EPI-NCSC hEPI-NCSC Pluripotency Bone Melanocyte