Faculty of Medicine and Health Sciences, Laboratory of Skin Neurobiology, University of Brest, Brest, France.
Exp Dermatol. 2012 Mar;21(3):195-200. doi: 10.1111/j.1600-0625.2011.01422.x.
Adult stem cells could be small sources of neurons or other cellular types for regenerative medicine and tissue engineering. Recently, pluripotent stem cells have been extracted from skin tissue, which opened a new accessible source for research. To routinely obtain a high yield of functional neurons from adult human skin stem cells with defined serum-free medium, stem cells from abdominal skin were cultured in serum-free medium. To differentiate them, we used a defined medium containing growth factors. Differentiated cells were identified using the following methods: (i) Oil-red-O staining for adipocytes, immunocytochemistry with antibodies recognising (ii) neurofilaments and PGP9.5 for neural differentiation, (iii) glial fibrillary acidic protein (GFAP) for glial differentiation, (iv) Ki-67 for proliferative cells, (v) FM1-43 staining to analyse vesicle trafficking in neuronal cells and (vi) a PCR array was used. Stem cells were floating in spheres and were maintained in culture for 4 months or more. They expressed nestin and Oct 4 and were proliferative. We induced specific differentiation into adipocytes, glial and neuronal cells. The yields of differentiated neurons were high and reproducible. They were maintained for long time (1 month) in the culture medium. Furthermore, these neurons incorporated FM1-43 dye, which indicates a potent acquisition of synaptic features in neurons. Stem cells from adult human skin could be valuable and reproducible tools/source to obtain high numbers of functional specific cellular types, such as neurons, for tissue engineering. In this work, the possibility to obtain a high yield of differentiated neurons, with the ability of endocytosis and vesicle cell trafficking, was shown.
成体干细胞可以作为再生医学和组织工程中神经元或其他细胞类型的小来源。最近，已从皮肤组织中提取出多能干细胞，这为研究开辟了新的可利用来源。为了从成人皮肤干细胞中常规获得高产率的功能性神经元，并使用无血清培养基，我们在无血清培养基中培养腹部皮肤的干细胞。为了使它们分化，我们使用含有生长因子的定义培养基。使用以下方法鉴定分化细胞：（i）油红-O 染色鉴定脂肪细胞，免疫细胞化学鉴定（ii）神经丝和 PGP9.5 用于神经分化，（iii）胶质纤维酸性蛋白（GFAP）用于神经胶质分化，（iv）Ki-67 用于增殖细胞，（v）FM1-43 染色分析神经元细胞中的囊泡运输，（vi）使用 PCR 阵列。干细胞在球体中漂浮，并在培养中维持 4 个月或更长时间。它们表达巢蛋白和 Oct 4，具有增殖能力。我们诱导其特异性分化为脂肪细胞、神经胶质细胞和神经元细胞。分化神经元的产量高且具有可重复性。它们在培养基中可以长时间（1 个月）维持。此外，这些神经元摄取 FM1-43 染料，这表明神经元获得了强大的突触特征。成人皮肤干细胞可以作为有价值的、可重复使用的工具/来源，用于获得大量的功能性特定细胞类型，如神经元，用于组织工程。在这项工作中，显示了获得高产率、具有内吞作用和囊泡细胞运输能力的分化神经元的可能性。
