Abstract
Since the first mammal was cloned, the idea of using this technique to help endangered species has aroused considerable interest. However, several issues limit this possibility, including the relatively low success rate at every stage of the cloning process, and the dearth of usable tissues from these rare animals. iPS cells have been produced from cells from a number of rare mammalian species and this is the method of choice for strategies to improve cloning efficiency and create new gametes by directed differentiation. Nevertheless information about other stem cell/progenitor capabilities of cells from endangered species could prove important for future conservation approaches and adds to the knowledge base about cellular material that can be extremely limited. Multipotent progenitor cells, termed skin-derived precursor (SKP) cells, can be isolated directly from mammalian skin dermis, and human cheek tissue has also been shown to be a good source of SKP-like cells. Recently we showed that structures identical to SKPs termed m-SKPs could be obtained from monolayer/ two dimensional (2D) skin fibroblast cultures. Here we aimed to isolate m-SKPs from cultured cells of three endangered species; giant panda (Ailuropoda melanoleuca); red panda (Ailurus fulgens); and Asiatic lion (Panthera leo persica). m-SKP-like spheres were formed from the giant panda buccal mucosa fibroblasts; whereas dermal fibroblast (DF) cells cultured from abdominal skin of the other two species were unable to generate spheres. Under specific differentiation culture conditions giant panda spheres expressed neural, Schwann, adipogenic and osteogenic cell markers. Furthermore, these buccal mucosa derived spheres were shown to maintain expression of SKP markers: nestin, versican, fibronectin, and P75 and switch on expression of the stem cell marker ABCG2. These results demonstrate that giant panda cheek skin can be a useful source of m-SKP multipotent progenitors. At present lack of sample numbers means that we can only postulate why we were unable to obtain m-SKPs from the lion and red panda cultures. However the giant panda observations point to the value of archiving cells from rare species, and the possibilities for later progenitor cell derivation.
Highlights
Skin-derived precursor (SKP) cells are a population of multipotent progenitor cells, which possess the ability to differentiate along various lineages including neural crest and mesenchymal progeny [1]
SKPs are conventionally obtained from freshly isolated tissue, but we have previously demonstrated that identical structures, termed m-SKPs, could be derived from monolayer human skin fibroblast cultures including those that had been previously cryopreserved [7]
Dermal fibroblast cells isolated from human skin from various body sites, including abdominal skin, have previously been shown to be capable of generating SKPs and m-SKPs [7,11], we were unable to generate mSKP-like spheroids when fibroblasts cultured from the abdominal tissue of the red panda and Asiatic lion were put into SKP medium (Fig 2)
Summary
Skin-derived precursor (SKP) cells are a population of multipotent progenitor cells, which possess the ability to differentiate along various lineages including neural crest and mesenchymal progeny [1]. Previous reports suggest that SKP numbers decline with age [5], a more recent report has shown that SKPs can efficiently be obtained from the cheek/chin tissue of aged individuals [6]. SKPs are conventionally obtained from freshly isolated tissue, but we have previously demonstrated that identical structures, termed m-SKPs, could be derived from monolayer human skin fibroblast cultures including those that had been previously cryopreserved [7]. Because the hair follicle dermal papilla has previously been identified as an enriched niche for SKPs [2,3,4], m-SKPs could not be derived from cultured human hair follicle dermal papilla cells in the same study. Suggesting that the body sites and tissues from which SKPs and m-SKPs can be produced are not entirely predictable
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