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

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.Electronic supplementary materialThe online version of this article (doi:10.1007/s12015-011-9255-5) contains supplementary material, which is available to authorized users.

Sox10+ adult stem cells contribute to biomaterial encapsulation and microvascularization

“…In recent years, Sox10 + stem cells have been reported to exist in multiple adult tissues, including inferior turbinate23, periodontal ligament24, hair follicle2526, mammary epithelium27, and bone marrow28. Sox10 could also promote tumor formation29.…”

The importance and use of stem cells in dermatology

“…rdzenia kręgowego. Naukowcom z Uniwersytetu w Newcastle udało się przekształcić komórki macierzyste skóry w osteocyty i melanocyty [13]. Od 2006 roku wykorzystywana jest metoda genetycznego przeprogramowania fibroblastów skóry in vitro tak, aby mogły cofnąć się do etapu niedojrzałego oraz niezróżnicowanego i były w stanie rozwijać się w różne linie komórkowe.…”

Is There a Role for Neural Crest Stem Cells in Periodontal Regeneration?

“…The presence of NCSC was demonstrated not only in the embryo but also in the adult tissues [8][9][10], and our group investigated the presence of NCSC in the adult PDL tissue using two immortalized clonal PDL cell lines with multipotency [63]. One of these, cell line 1-11, had the capacity to differentiate into osteoblasts and adipocyte, whereas the other clone, cell line 1-17, could differentiate into osteoblasts, chondrocytes, adipocytes, and neural cell.…”

“…Recent reports have demonstrated that NCSC are present not only in the neural crest in the embryo but also in their target tissues in fetal and adult tissues; human epidermal NCSC are multipotent remnants in a postnatal location. For example, NCSC can be found in the bulge of hair follicles where they persist postnatally and into adulthood [8][9][10]. In addition, many reports have identified the presence of stem cells in NCSC-derived dental tissues such as dental follicle, apical papilla, pulp, exfoliated deciduous teeth, and PDL, and numerous in vitro and in vivo studies [16-22, 24-45, 49-69].…”