EGF-FGF2 stimulates the proliferation and improves the neuronal commitment of mouse epidermal neural crest stem cells (EPI-NCSCs)
EGF-FGF2 stimulates the proliferation and improves the neuronal commitment of mouse epidermal neural crest stem cells (EPI-NCSCs)
复制标题
DOI：
10.1016/j.yexcr.2014.05.020
复制
发表时间：
2014-09-10
影响因子：
3.7
通讯作者：
Trentin, Andrea Goncalves
中科院分区：
文献类型：
--
作者：
Bressan, Raul Bardini;Melo, Fernanda Rosene;Trentin, Andrea Goncalves
表皮神经嵴干细胞（EPI-NCSC）位于毛囊的隆突处，在细胞治疗、药物筛选和组织工程等方面具有广泛的应用前景。正如所建议的胚胎神经嵴（NC）在成人位置的残余物，EPI-NCSC能够产生各种各样的细胞类型，并易于通过微创手术获得。由于表皮生长因子（EGF）和2型成纤维细胞生长因子（FGF（2））的组合是促有丝分裂的，并促进各种干细胞群的神经元定型，我们研究了其在小鼠EPI-NCSC的增殖和神经元潜能中的作用。通过使用公认的隆须卵泡培养方案，我们能够分离EPI-NCSC的群体，其特征在于NC细胞的迁移潜力、细胞形态和表型标记物的表达。EPI-NCSCs表达神经元、神经胶质和平滑肌标志物，并呈现NC样成纤维细胞形态。然而，EGF和FGF（2）联合治疗增加了它们的增殖率，促进了神经元样形态的获得，伴随着神经细胞骨架蛋白β III-微管蛋白和巢蛋白的重组，以及泛神经元标记β III-微管蛋白的上调和未分化NC、神经胶质和平滑肌细胞标记的下调。此外，治疗增强了EPI-NCSC对神经源性刺激的反应，如通过GAP 43的诱导和神经元样细胞中Mash-1（两者均为神经元特异性蛋白）的表达增加所证明的。总之，结果表明EGF FGF（2）的组合刺激EPI-NCSC的增殖并改善其神经元潜能，类似于胚胎NC细胞、ES细胞和中枢神经系统的神经祖细胞/干细胞，并突出了在神经元分化方案中使用EGF-FGF（2）的优势。(C)2014爱思唯尔公司All rights reserved.
Epidermal neural crest stem cells (EPI-NCSCs), which reside in the bulge of hair follicles, are attractive candidates for several applications in cell therapy, drug screening and tissue engineering. As suggested remnants of the embryonic neural crest (NC) in an adult location, EPI-NCSCs are able to generate a wide variety of cell types and are readily accessible by a minimally invasive procedure. Since the combination of epidermal growth factor (EGF) and fibroblast growth factor type 2 (FGF(2)) is mitogenic and promotes the neuronal commitment of various stem cell populations, we examined its effects in the proliferation and neuronal potential of mouse EPI-NCSCs. By using a recognized culture protocol of bulge whiskers follicles, we were able to isolate a population of EPI-NCSCs, characterized by the migratory potential, cell morphology and expression of phenotypic markers of NC cells. EPI-NCSCs expressed neuronal, glial and smooth muscle markers and exhibited the NC-like fibroblastic morphology. The treatment with the combination EGF and FGF(2), however, increased their proliferation rate and promoted the acquisition of a neuronal-like morphology accompanied by reorganization of neural cytoskeletal proteins beta III-tubulin and nestin, as well as upregulation of the pan neuronal marker beta III-tubulin and down regulation of the undifferentiated NC, glial and smooth muscle cell markers. Moreover, the treatment enhanced the response of EPI-NCSCs to neurogenic stimulation, as evidenced by induction of GAP43, and increased expression of Mash-1 in neuron-like cell, both neuronal-specific proteins. Together, the results suggest that the combination of EGF FGF(2) stimulates the proliferation and improves the neuronal potential of EPI-NCSCs similarly to embryonic NC cells, ES cells and neural progenitor/stem cells of the central nervous system and highlights the advantage of using EGF-FGF(2) in neuronal differentiation protocols. (C) 2014 Elsevier Inc. All rights reserved.