Liu Fang, Hoffman Robert M
Department of Anatomy, Regenerative Medicine Center, Second Military Medical University, Shanghai, China.
AntiCancer, Inc., San Diego, CA, USA.
Methods Mol Biol. 2018;1760:145-162. doi: 10.1007/978-1-4939-7745-1_15.
The stem cell marker, nestin, is expressed in the hair follicle, both in cells in the bulge area (BA) and the dermal papilla (DP). Nestin-expressing hair follicle-associated-pluripotent (HAP) stem cells of both the BA and DP have been previously shown to be able to form neurons, heart muscle cells, and other non-follicle cell types. The ability of the nestin-expressing HAP stem cells from the BA and DP to repair spinal cord injury was compared. Nestin-expressing HAP stem cells from both the BA and DP grew very well on Gelfoam. The HAP stem cells attached to the Gelfoam within 1 h. They grew along the grids of the Gelfoam during the first 2 or 3 days. Later they spread into the Gelfoam. After transplantation of Gelfoam cultures of nestin-expressing BA or DP HAP stem cells into the injured spinal cord (including the Gelfoam) nestin-expressing BA and DP cells were observed to be viable over 100 days post-surgery. Hematoxylin and eosin (H&E) staining showed connections between the transplanted cells and the host spine tissue. Immunohistochemistry showed many Tuj1-, Isl 1/2, and EN1-positive cells and nerve fibers in the transplanted area of the spinal cord after BA Gelfoam or DP Gelfoam cultures were transplanted to the spine. The spinal cord of mice was injured to effect hind-limb paralysis. Twenty-eight days after transplantation with BA or DP HAP stem cells on Gelfoam to the injured area of the spine, the mice recovered normal locomotion.
干细胞标志物巢蛋白在毛囊中表达，在隆突区（BA）和真皮乳头（DP）的细胞中均有表达。先前已表明，BA和DP中表达巢蛋白的毛囊相关多能（HAP）干细胞能够形成神经元、心肌细胞和其他非毛囊细胞类型。比较了BA和DP中表达巢蛋白的HAP干细胞修复脊髓损伤的能力。BA和DP中表达巢蛋白的HAP干细胞在明胶海绵上生长良好。HAP干细胞在1小时内附着于明胶海绵。在最初的2或3天内，它们沿着明胶海绵的网格生长。随后它们扩散到明胶海绵中。将表达巢蛋白的BA或DP HAP干细胞的明胶海绵培养物移植到受损脊髓（包括明胶海绵）后，观察到表达巢蛋白的BA和DP细胞在手术后100天以上仍存活。苏木精和伊红（H&E）染色显示移植细胞与宿主脊柱组织之间有连接。免疫组织化学显示，将BA明胶海绵或DP明胶海绵培养物移植到脊柱后，脊髓移植区域有许多Tuj1、Isl 1/2和EN1阳性细胞及神经纤维。小鼠脊髓受损导致后肢麻痹。将BA或DP HAP干细胞与明胶海绵一起移植到脊柱损伤区域28天后，小鼠恢复了正常运动。
