Human Schwann Cells Retain Essential Phenotype Characteristics After Immortalization
Human Schwann Cells Retain Essential Phenotype Characteristics After Immortalization
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DOI：
10.1089/scd.2010.0513
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发表时间：
2012-02-10
影响因子：
4
通讯作者：
Hoeke, Ahmet
中科院分区：
文献类型：
--
作者：
Lehmann, Helmar C.;Chen, Weiran;Hoeke, Ahmet
雪旺细胞（Schwann cells，SC）在周围神经疾病的发病机制中起着重要作用，是一种潜在的治疗靶点。然而，将原代人SC（hSC）用于体外模型是有限的，因为这些细胞难以在普通细胞培养条件下制备并保持高产率和纯度。为了克服这一障碍，我们使用SV 40大T抗原和人端粒酶逆转录酶表达载体永生化原代人胎儿干细胞。经过克隆，选择和纯化，我们评估了几个永生SC线的表达细胞外基质（ECM）分子和髓鞘化胚胎大鼠感觉轴突的能力。此外，我们建立了一个基因表达谱，并探讨了他们的敏感性，在一个简单的体外测定氧化应激。通过免疫细胞化学、微阵列和定量逆转录聚合酶链反应测定，永生化hSC克隆表达常见的胶质细胞标志物和多种生长因子、受体和ECM分子。在神经元-SC共培养中，这些细胞能够使大鼠背根神经节神经元髓鞘化，尽管它们的有效性低于原代大鼠SC。在毒性测定中，永生化hSC仍然对H2 O2诱导的氧化应激敏感。这项研究表明，使用特定的永生化技术，有可能建立保留典型原代hSC特征的hSC系。这些细胞对于药物筛选和针对涉及SC的疾病机制的研究特别有用。
Schwann cells (SCs) play an important role in the pathogenesis of peripheral nerve diseases and represent a potential target for development of therapies. However, use of primary human SCs (hSCs) for in vitro models is limited because these cells are difficult to prepare and maintain in high yield and purity under common cell culture conditions. To circumvent this obstacle, we immortalized primary human fetal SCs using the SV40 large T-antigen and human telomerase reverse transcriptase expression vectors. After cloning, selection, and purification, we evaluated several immortalized SC lines for their ability to express extracellular matrix (ECM) molecules and myelinate embryonic rat sensory axons. In addition, we established a gene expression profile and explored their sensitivity to oxidative stress in a simple in vitro assay. Immortalized hSC clones expressed common glial markers and a broad variety of growth factors, receptors, and ECM molecules as determined by immunocytochemistry, microarray, and quantitative reverse transcription polymerase chain reaction. In neuron-SC co-cultures, these cells were able to myelinate rat dorsal root ganglia neurons, although their effectiveness was lower in comparison to primary rat SCs. In toxicity assays, immortalized hSCs remain susceptible to oxidative stress induced by H2O2. This study shows that, using specific immortalization techniques, it is possible to establish hSC lines that retain characteristics of typical primary hSCs. These cells are particularly useful for drug screening and studies aimed at disease mechanisms involving SCs.