Establishment and characterization of spontaneously immortalized Schwann cells from murine model of globoid cell leukodystrophy (twitcher)
Establishment and characterization of spontaneously immortalized Schwann cells from murine model of globoid cell leukodystrophy (twitcher)
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DOI：
10.1002/jnr.10247
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发表时间：
2002-06-01
影响因子：
4.2
通讯作者：
Eto, Y
中科院分区：
文献类型：
--
作者：
Shen, JS;Watabe, K;Eto, Y
Twitcher小鼠是人类脑白质细胞营养不良症(GLD；Krabbe病)的小鼠模型，由半乳糖神经酰胺酶(GALC)活性的遗传缺陷引起。在抽动小鼠和GLD患者的髓鞘形成细胞(少突胶质细胞和雪旺细胞)中，细胞毒性代谢物半乳糖鞘氨醇(神经氨酸)的积累被认为会导致这些细胞的功能障碍，并随后导致中枢和外周神经系统的脱髓鞘。为了进一步研究GLD的细胞病理机制，我们建立了Twitcher小鼠自发永生化的雪旺细胞系。取3周龄Twitcher小鼠背根节和连续外周神经来源的雪旺细胞长期培养6个月，进一步扩增并鉴定自发形成的集落。其中一个细胞系，命名为TwS1，显示出明显的雪旺细胞表型，每周传代两次，并保持了10个月以上，没有表型变化。TwS1细胞在GALC基因组中发生了无义突变，GALC活性显著降低，精神肽水平显著升高。在超微结构下，TwS1细胞内可见多种胞质内含物。当TwS1细胞被编码GALC的逆转录病毒载体感染时，GALC活性显著增加，精神素水平显著降低。这些永生化的雪旺细胞可用于GLD神经系统损害的研究。(C)2002年Wiley-Liss，Inc.
The twitcher mouse is a murine model of human globoid cell leukodystrophy (GLD; Krabbe disease) caused by a genetic defect in the activity of galactosylceramidase (GALC). An accumulation of cytotoxic metabolite, galactosylsphingosine (psychosine), in myelin forming cells (oligodendrocytes and Schwann cells) of the twitcher mouse as well as patients with GLD has been suggested to cause dysfunction of these cells and subsequent demyelination in the central and peripheral nervous system. To investigate further the cellular pathomechanism of GLD, we established spontaneously immortalized Schwann cell lines from the twitcher mouse. Long-term cultures of Schwann cells derived from dorsal root ganglia and consecutive peripheral nerves of 3-week-old twitcher mice were maintained for 6 months, and spontaneously developed colonies were expanded further and characterized. One of the cell lines, designated TwS1, showed distinct Schwann cell phenotypes, was passaged twice a week and maintained for over 10 months without phenotypic alterations. The TwS1 cells had a nonsense mutation in the GALC genome, and showed markedly reduced GALC activity and elevated psychosine levels. Ultrastructurally, varieties of cytoplasmic inclusions were demonstrated in TwS1 cells. When TwS1 cells were infected with a retrovirus vector encoding GALC, GALC activity was markedly increased and psychosine levels were significantly decreased. These immortalized Schwann cells can be useful in studies on the nervous system lesions in GLD. (C) 2002 Wiley-Liss, Inc.