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

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

Impact of climate change and loss of habitat on sirenians

Although the impacts of climate change on the welfare of individual manatees and dugongs are still uncertain, the effects are likely to be through indirect interactions between meteorological and biotic factors and the human responses to climate change. We divided the potential impacts into (1) those that will potentially affect sirenians directly including temperature increases, sea-level rise, increased intensity of extreme weather events and changes in rainfall patterns and (2) indirect impacts that are likely to cause harm through habitat loss and change and the increase in the likelihood of harmful algal blooms and disease outbreaks. The habitat modification accompanying sea-level rise is likely to decrease the welfare of sirenians including increased mortality. Many species of tropical seagrasses live close to their thermal limits and will have to up-regulate their stress-response systems to tolerate the sublethal temperature increases caused by climate change. The capacity of seagrass species to evoke such responses is uncertain, as are the effects of elevated carbon dioxide on such acclimation responses. The increase in the intensity of extreme weather events associated with climate change is likely to decrease the welfare of sirenians through increased mortality from strandings, as well as habitat loss and change. These effects are likely to increase the exposure of sirenians to disease and their vulnerability to predators, including human hunters. Climate-related hazards will also exacerbate other stressors, especially for people living in poverty. Thus the risks to sirenians from climate change are likely to be greatest for small populations of dugongs and manatees occurring in low-income countries. The African manatee will be particularly vulnerable because of the high levels of human poverty throughout most of its range resulting in competition for resources, including protein from manatee meat