A two-compartment organotypic model of mammalian peripheral nerve repair.
A two-compartment organotypic model of mammalian peripheral nerve repair.
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DOI：
10.1016/j.jneumeth.2014.05.005
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发表时间：
2014-07-30
影响因子：
3
通讯作者：
Brushart, Thomas M.
中科院分区：
文献类型：
--
作者：
Siddique, Rezina;Vyas, Alka;Thakor, Nitish;Brushart, Thomas M.
横断神经远端残端的雪旺细胞可上调生长因子，以支持特定形态的再生。然而，目前尚不清楚这些细胞中哪些优先支持运动轴突再生。识别这些因素将需要一个模型，该模型可以分离生长因子对生长轴突的影响，同时再现周围神经的复杂三维结构。由两个腔室组成的PDMS底座顶部覆盖着一层胶原膜，它支撑着一个腔室上方的脊髓横截面。本部分中的荧光运动神经元对周围神经的一段进行再神经支配，该神经通过防水屏障引导轴突到第二隔室，在那里进行神经修复。运动神经元可以在几周内保持健康。他们通过防水屏障投射的轴突在横断后存活下来，并大量穿过神经修复，重新支配额外的神经节。通过染料渗漏试验确认两个隔室的流体隔离，并通过仅对暴露于示踪剂的运动神经元进行逆行标记并将毒素效应限制在单个隔室来测试系统的生理完整性。神经修复不能在单层细胞培养中建模。我们之前的器官模型准确地模拟了神经修复，但不允许个人控制运动神经元和生长锥环境。该模型分离了对生长轴突的治疗效果，同时再现了周围神经的复杂三维结构。此外，它还便于组织的外科操作和高分辨率成像。
Schwann cells in the distal stump of transected nerve upregulate growth factors that support regeneration on a modality-specific basis. It is unclear, however, which of these preferentially support motor axon regeneration. Identification of these factors will require a model that can isolate growth factor effects to growing axons while reproducing the complex three-dimensional structure of peripheral nerve. A two-compartment PDMS base is topped by a collagen-coated membrane that supports a spinal cord cross-section above one compartment. Fluorescent motoneurons in this section reinnervate a segment of peripheral nerve that directs axons through a watertight barrier to the second compartment, where nerve repair is performed. Motoneurons remain healthy for several weeks. The axons they project through the water-tight barrier survive transection and cross a nerve repair in substantial numbers to reinnervate an additional nerve segment. Fluidic isolation of the two compartments was confirmed with a dye leakage test, and the physiologic integrity of the system was tested by retrograde labeling of only those motor neurons to which tracer was exposed and by limitation of toxin effects to a single compartment. Nerve repair cannot be modeled in monolayer cell culture. Our previous organotypic model accurately modeled nerve repair, but did not allow individual control of motoneuron and growth cone environments. This model isolates treatment effects to growing axons while reproducing the complex three-dimensional structure of peripheral nerve. Additionally, it facilitates surgical manipulation of tissues and high-resolution imaging.
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影响因子：
6.1
作者：
Hosmane, Suneil;Yang, In Hong;Venkatesan, Arun
通讯作者：
Venkatesan, Arun
影响因子：
2.7
作者：
CAMPENOT, RB
通讯作者：
CAMPENOT, RB
影响因子：
5.3
作者：
Yang, In Hong;Siddique, Rezina;Hosmane, Suneil;Thakor, Nitish;Hoeke, Ahmet
通讯作者：
Hoeke, Ahmet
影响因子：
2.9
作者：
IDE, C;TOHYAMA, K;ONODERA, S
通讯作者：
ONODERA, S
DOI：
10.1002/(sici)1097-4695(199710)33:4
发表时间：
1997-10-01
期刊：
JOURNAL OF NEUROBIOLOGY
影响因子：
--
作者：
Kimpinski, K;Campenot, RB;Mearow, K
通讯作者：
Mearow, K