aFGF promotes axonal growth in rat spinal cord organotypic slice co-cultures
aFGF promotes axonal growth in rat spinal cord organotypic slice co-cultures
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DOI：
10.1089/089771502753594927
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发表时间：
2002-03-01
影响因子：
4.2
通讯作者：
Robertson, RT
中科院分区：
文献类型：
--
作者：
Lee, YS;Baratta, J;Robertson, RT
本研究开发了切片培养模型系统来研究脊髓损伤后的轴突再生。该模型在酸性成纤维细胞生长因子（aFGF）和周围神经节段在脊髓片之间轴突生长中的作用的研究中进行了测试。收集 P15-P18 Sprague-Dawley 大鼠脊髓的横切片用于器官切片培养。第一组由培养期间相互接触的两片脊髓组成。第二组由两片相距 3 毫米并由两段肋间神经连接的切片组成。第三组由用于研究神经元存活的单片组成。每组的一些培养物的培养基中都含有 aFGF。某些培养物的培养基中包含溴脱氧尿苷 (BrdU)。结果显示了三个主要发现。首先，神经丝阳性细胞计数表明，aFGF 处理显着增加了培养物中存活神经元的数量。其次，神经丝免疫染色和 DiI 示踪表明轴突穿过两段脊髓之间的连接处或穿过肋间神经段生长，并且这些轴突仅在经过 aFGF 处理的培养物中可见。第三，很少有细胞被神经丝和 BrdU 双重染色，并且这些细胞仅在 aFGF 处理后才被发现。这些结果表明（1）器官切片培养物提供了研究脊髓损伤再生的有用模型，（2）aFGF 拯救神经元并促进这些培养物中的轴突生长，（3）肋间神经节段促进脊髓切片之间的轴突生长。
This study developed a slice culture model system to study axonal regeneration after spinal cord injury. This model was tested in studies of the roles of acidic fibroblast growth factor (aFGF) and peripheral nerve segments in axonal growth between pieces of spinal cord. Transverse sections of P15-P18 Sprague-Dawley rat spinal cord were collected for organotypic slice cultures. Group I consisted of two slices of spinal cord in contact with each other during the culture period. Group II consisted of two slices that were separated by 3 mm and connected by two segments of intercostal nerves. Group III consisted of single slices for studies of neuron survival. Some cultures from each group included aFGF in the culture medium. Bromodeoxyuridine (BrdU) was included in the medium for some cultures. The results showed three principal findings. First, counts of neurofilament-positive cells demonstrated that treatment with aFGF significantly increased the number of surviving neurons in culture. Second, neurofilament immunostaining and DiI tracing demonstrated axons crossing the junction between the two pieces of spinal cord or growing through the intercostal nerve segments, and these axons were seen only in cultures with aFGF treatment. Third, few cells were double stained for neurofilament and BrdU, and these were found only with aFGF treatment. These results demonstrate that (1) organotypic slice cultures present a useful model to study regeneration from spinal cord injury, (2) aFGF rescues neurons and promotes axonal growth in these cultures, and (3) segments of intercostal nerves promote axon growth between slices of spinal cord.