Division of Genetics and Development, Krembil Neuroscience Centre, Toronto Western Research Institute, University Health Network, Toronto, Ontario, Canada.
Stem Cells Dev. 2012 Feb 10;21(3):432-47. doi: 10.1089/scd.2011.0242. Epub 2011 Dec 5.
In cellular transplantation strategies for repairing the injured central nervous system, interactions between transplanted neural precursor cells (NPCs) and host tissue remain incompletely understood. Although trophins may contribute to the benefits observed, little research has explored this possibility. Candidate trophic factors were identified, and primers were designed for these genes. Template RNA was isolated from 3 NPC sources, and also from bone marrow stromal cells (BMSCs) and embryonic fibroblasts as comparative controls. Quantitative polymerase chain reaction was performed to determine the effect of cell source, passaging, cellular differentiation, and environmental changes on trophin factor expression in NPCs. Results were analyzed with multivariate statistical analyses. NPCs, BMSCs, and fibroblasts each expressed trophic factors in unique patterns. Trophic factor expression was similar among NPCs whether harvested from rat or mouse, brain or spinal cord, or their time in culture. The expression of neurotrophin NT-3, NT-4/5, glial-derived neurotrophic factor, and insulin-like growth factor-1 decreased with time in culture. Induced differentiation of NPCs led to a marked and statistically significant increase in the expression of trophic factors. Culture conditions and environmental changes were also associated with significant changes in trophin expression. These results suggest that trophins could contribute to the benefits associated with transplantation of NPCs as well as BMSCs. Trophic factor expression changes with NPC differentiation and environmental conditions, which could have important implications with regard to their behavior after in vivo transplantation.
在修复中枢神经系统损伤的细胞移植策略中,移植的神经前体细胞(NPCs)与宿主组织之间的相互作用仍不完全清楚。尽管神经营养因子可能有助于观察到的益处,但很少有研究探讨这种可能性。确定了候选的营养因子,并为这些基因设计了引物。从 3 种 NPC 来源中分离出模板 RNA,并从骨髓基质细胞(BMSCs)和胚胎成纤维细胞中分离出作为比较对照的 RNA。进行定量聚合酶链反应以确定细胞来源、传代、细胞分化和环境变化对 NPC 中营养因子表达的影响。使用多变量统计分析对结果进行分析。NPC、BMSCs 和成纤维细胞各自以独特的模式表达营养因子。无论从大鼠还是小鼠、脑或脊髓中提取 NPC,或培养时间长短,NPC 中的营养因子表达相似。神经营养因子 NT-3、NT-4/5、胶质源性神经营养因子和胰岛素样生长因子-1 的表达随培养时间的延长而减少。NPC 的诱导分化导致营养因子的表达显著增加。培养条件和环境变化也与营养因子表达的显著变化有关。这些结果表明,神经营养因子可能有助于 NPC 以及 BMSCs 移植相关的益处。营养因子的表达随 NPC 分化和环境条件的变化而变化,这可能对其体内移植后的行为有重要影响。