BDNF, NT-3, and NGF released from transplanted neural progenitor cells promote corticospinal axon growth in organotypic cocultures
BDNF, NT-3, and NGF released from transplanted neural progenitor cells promote corticospinal axon growth in organotypic cocultures
复制标题
DOI：
10.1097/brs.0b013e318059afab
复制
发表时间：
2007-05-20
期刊：
影响因子：
3
通讯作者：
Ochi, Mitsuo
中科院分区：
文献类型：
--
作者：
Kamei, Naosuke;Tanaka, Nobuhiro;Ochi, Mitsuo
研究设计。目的：应用器官型切片培养技术对脊髓损伤进行实验研究。目的：探讨神经祖细胞移植损伤脊髓后皮质-脊髓轴突再生的机制。背景资料摘要。npc移植后中枢神经系统再生的几种机制已被提出；然而，确切的机制还不清楚。在此之前，我们利用新生大鼠大脑皮层和脊髓的独特器官型共培养系统，证明了移植的npc分泌体液因子，进而促进皮质-脊髓轴突生长。培养的NPCs在分化前后分别用抗脑源性神经营养因子（BDNF）、神经营养因子(NT)-3、神经生长因子（NGF）和睫状神经营养因子（CNTF）的抗体进行免疫染色。为了定量评估皮质脊髓轴突的生长，我们使用了器官型共培养系统。从新生大鼠获得的解剖脑皮层和脊髓彼此排列，并在膜上培养。npc被移植到共培养物上。此外，在共培养中加入抗BDNF、NT-3、NGF或CNTF的中和抗体。用dil顺行轴突示踪法定量评价轴突从大脑皮层向脊髓的生长。培养的NPCs在分化前后均被BDNF、NT3、NGF和cttnf抗体免疫染色呈阳性。npc移植促进了轴突从大脑皮层向脊髓的生长。添加抗BDNF、NT-3和NGF的中和抗体可显著抑制NPCs促进的轴突生长，但不抑制cntf。移植的npc分泌的神经营养因子BDNF、NT-3和NGF参与了npc移植后皮质脊髓轴突生长的促进。
Study Design. Experimental study of spinal cord injury using an organotypic slice culture.Objective. To clarify the mechanism of corticospinal axon regeneration following transplantation of neural progenitor cells (NPCs) in the injured spinal cord.Summary of Background Data. Several mechanisms underlying central nervous system regeneration after transplantation of NPCs have been proposed; however, the precise mechanism has not been clarified. Previously, we demonstrated that transplanted NPCs secreted humoral factors that in turn promoted corticospinal axon growth using the unique organotypic coculture system involving brain cortex and spinal cord from neonatal rats.Methods. Cultured NPCs were immunostained with antibodies against neurotrophic factors including brain-derived neurotrophic factor (BDNF), neurotrophin (NT)-3, nerve growth factor (NGF), and ciliary neurotrophic factor (CNTF) both before and after differentiation. To evaluate corticospinal axon growth quantitatively, we used the organotypic coculture system. The dissected brain cortex and spinal cord obtained from neonatal rats were aligned next to each other and cultured on a membrane. NPCs were transplanted onto the cocultures. Furthermore, neutralizing antibodies against BDNF, NT-3, NGF, or CNTF were added to the cocultures. Axon growth from the brain cortex into the spinal cord was assessed quantitatively using anterograde axon tracing with Dil.Results. The cultured NPCs were positively immunostained by antibodies against BDNF, NT3, NGF, and CTNF both before and after differentiation. Transplantation of NPCs promoted axon growth from the brain cortex into the spinal cord. The axon growth promoted by NPCs was significantly suppressed by the addition of neutralizing antibodies against BDNF, NT-3, and NGF but not CNTF.Conclusion. The neurotrophic factors, BDNF, NT-3, and NGF, secreted by transplanted NPCs, were involved in the promotion of corticospinal axon growth after transplantation of NPCs.