Dept. of Neurosciences, University of California, San Diego, La Jolla, CA, USA.
Dept. of Neurosciences, University of California, San Diego, La Jolla, CA, USA; Veterans Administration San Diego Healthcare System, San Diego, CA, USA.
Exp Neurol. 2017 May;291:87-97. doi: 10.1016/j.expneurol.2017.02.007. Epub 2017 Feb 9.
Previously we utilized fibrin matrices and a cocktail of nine growth factors and a cell death inhibitor to promote survival and fill of neural stem cell (NSC) or neural progenitor cell (NPC) grafts to sites of spinal cord injury (SCI). In the current study, we examined whether the number of growth factors in a supportive matrix could be reduced to a more clinically practical number while retaining extensive NPC survival and fill of a spinal cord lesion site. NPCs derived from embryonic day 14 Fischer 344 rat spinal cords expressing green fluorescent protein (GFP) were embedded in fibrin matrices containing a defined growth factor cocktail: one to four factors were tested among nine different groups. Grafts were placed into C5 lateral hemisection lesion sites two weeks post-injury, and graft survival and fill was assessed two weeks later. We found that a four growth factor cocktail consisting of brain-derived neurotrophic factor (BDNF), basic-fibroblastic growth factor (bFGF), vascular endothelial growth factor (VEGF), and MDL28170, a cell death inhibitor, resulted in consistent graft survival, neuronal differentiation, and fill of the lesion site. Extensive stem cell-derived axonal outgrowth from the lesion site occurred, consistent with previous reports. Fewer than four growth factors resulted in suboptimal NPC fill of the lesion site. Collectively, these findings indicate that a simplified, four-component cocktail can support neural progenitor cell engraftment to a spinal cord lesion site to the same extent as a 10-component cocktail.
先前，我们利用纤维蛋白基质和包含九种生长因子和一种细胞死亡抑制剂的鸡尾酒来促进神经干细胞（NSC）或神经前体细胞（NPC）移植到脊髓损伤（SCI）部位的存活和填充。在本研究中，我们考察了在保留大量 NPC 存活和填充脊髓损伤部位的情况下，支持基质中的生长因子数量是否可以减少到更符合临床实际的数量。从胚胎 14 天 Fischer 344 大鼠脊髓中分离出表达绿色荧光蛋白（GFP）的 NPC，将其嵌入含有特定生长因子鸡尾酒的纤维蛋白基质中：在 9 个不同组中测试了一个到四个因子。在损伤后两周将移植物置于 C5 侧半切损伤部位，两周后评估移植物的存活和填充情况。我们发现，由脑源性神经营养因子（BDNF）、碱性成纤维细胞生长因子（bFGF）、血管内皮生长因子（VEGF）和细胞死亡抑制剂 MDL28170 组成的四因子鸡尾酒可导致一致的移植存活、神经元分化和损伤部位的填充。从损伤部位发生了广泛的干细胞源性轴突延伸，与之前的报告一致。少于四个因子会导致 NPC 对损伤部位的填充不足。总的来说，这些发现表明，简化的四因子鸡尾酒可以支持神经前体细胞移植到脊髓损伤部位，其效果与十因子鸡尾酒相同。
