A novel method for isolating Schwann cells using the extracellular domain of Necl1.

A novel method for isolating Schwann cells using the extracellular domain of Necl1.
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DOI:
10.1002/jnr.21985
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发表时间:
2009-11-15
影响因子:
4.2
通讯作者:
Peles, Elior
Peles, Elior
中科院分区:
医学3区
文献类型:
--
作者:
Spiegel, Ivo;Peles, Elior

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雪旺细胞与drg -神经元髓鞘共培养是探索轴突-雪旺细胞相互作用分子机制的有力实验系统。分离出具有成髓能力的雪旺细胞的纯群体是这个实验系统的先决条件。我们在这里描述了一种使用特异性亲和试剂(Necl1-Fc)的基于facs的雪旺细胞分离的方案,并将这些分离的细胞用于髓鞘共培养。髓鞘共培养系统的一个优点是,雪旺细胞和神经元可以在共培养之前进行基因操作。我们进一步表明,我们的方法可以在一个纯化步骤中分离出病毒转导的雪旺细胞。通过Necl1-Fc在facs基础上分离具有髓鞘形成能力的雪旺细胞,并将这些细胞用于髓鞘形成共培养,这将极大地促进未来旨在描述轴突-雪旺细胞相互作用和髓鞘形成的分子机制的研究。
Myelinating co-cultures of Schwann cells and DRG-neurons are a powerful experimental system to probe the molecular mechanisms of axon-Schwann cell interaction. The isolation of a pure population of myelination-competent Schwann cells is a prerequisite for this experimental system. We describe here a protocol for a FACS-based isolation of Schwann cells utilizing a specific affinity-reagent (Necl1-Fc), and the use of these isolated cells in myelinating co-cultures. An advantage of the myelinating co-culture system is that Schwann cells and the neurons can be genetically manipulated before they are co-cultured. We further show that our method allows the isolation of virally transduced Schwann cells in a single purification step. This protocol for the FACS-based isolation of myelination-competent Schwann cells by Necl1-Fc and the use of these cells in myelinating co-cultures should significantly facilitate future studies aimed at the delineation of the molecular mechanisms axon-Schwann cell interactions and myelination.
DOI: 10.1083/jcb.32.2.439
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