Hoffman Robert M, Mii Sumiyuki, Duong Jennifer, Amoh Yasuyuki
AntiCancer Inc., San Diego, CA, USA.
Department of Surgery, UCSD, San Diego, CA, USA.
Methods Mol Biol. 2018;1760:163-186. doi: 10.1007/978-1-4939-7745-1_16.
Nestin-expressing hair follicle-associated pluripotent (HAP) stem cells reside mainly in the bulge area (BA) of the hair follicle but also in the dermal papilla (DP). The BA appears to be origin of HAP stem cells. Long-term Gelfoam histoculture was established of whiskers isolated from transgenic mice, in which there is nestin-driven green fluorescent protein (ND-GFP). HAP stem cells trafficked from the BA toward the DP area and extensively grew out onto Gelfoam forming nerve-like structures. These fibers express the neuron marker β-III tubulin-positive fibers and consisted of ND-GFP-expressing cells and extended up to 500 mm from the whisker nerve stump in Gelfoam histoculture. The growing fibers had growth cones on their tips expressing F-actin indicating that the fibers were growing axons. HAP stem cell proliferation resulted in elongation of the follicle nerve and interaction with other nerves in 3D Gelfoam histoculture, including the sciatic nerve, trigeminal nerve, and trigeminal nerve ganglion.
表达巢蛋白的毛囊相关多能（HAP）干细胞主要位于毛囊的隆突区（BA），但也存在于真皮乳头（DP）中。隆突区似乎是HAP干细胞的起源地。利用从转基因小鼠分离的须建立了长期的明胶海绵组织培养，这些转基因小鼠中存在巢蛋白驱动的绿色荧光蛋白（ND-GFP）。HAP干细胞从隆突区向真皮乳头区域迁移，并在明胶海绵上广泛生长，形成神经样结构。这些纤维表达神经元标志物β-III微管蛋白阳性纤维，由表达ND-GFP的细胞组成，在明胶海绵组织培养中从须神经残端延伸长达500毫米。生长的纤维尖端有表达F-肌动蛋白的生长锥，表明这些纤维是正在生长的轴突。在三维明胶海绵组织培养中，HAP干细胞增殖导致毛囊神经延长并与其他神经相互作用，包括坐骨神经、三叉神经和三叉神经节。
