Isolation and functional characterization of Schwann cells derived from adult peripheral nerve
Isolation and functional characterization of Schwann cells derived from adult peripheral nerve
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DOI：
10.1523/jneurosci.11-08-02433.1991
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发表时间：
1991-08
期刊：
影响因子：
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通讯作者：
T. K. Morrissey;N. Kleitman;R. Bunge
中科院分区：
文献类型：
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作者：
T. K. Morrissey;N. Kleitman;R. Bunge
为了促进自体移植技术的发展，以测试雪旺细胞（ScC）植入治疗神经系统损伤的能力，我们开发了一种从成人周围神经获取大量基本上纯的 ScC 群体的方法。通过允许周围神经干的小外植体在体外经历轴突和髓磷脂分解，而不是在收获后立即解离神经，我们能够（1）去除几乎所有成纤维细胞的外植体，并且（2）利用内在的 ScC 对周围神经变性的有丝分裂反应。在这里，我们描述了一种从成年大鼠坐骨神经中产生高达 98% 纯 ScC 群体的方法（基于细胞体和核形态、S100 染色以及神经突上解离细胞的行为），细胞产量大于 2 x 10(4) 细胞/毫克起始神经重量。纯化技术成功应用于人体组织；人膈神经产生了 98% 纯的 ScC 群体，细胞产量为 2 x 10(4) 细胞/毫克初始神经重量。与新生儿来源的 ScC 类似，成年大鼠细胞可以与背根神经节 (DRG) 神经元共培养，或在神经胶质生长因子和毛喉素存在下单独培养。细胞在 DRG 神经元上无限增殖，或者在化学有丝分裂剂下长达 10 周，在去除促有丝分裂刺激后恢复静止状态。扩增的成年大鼠 ScC 保留了功能能力，其将 DRG 神经突髓鞘化并支持胚胎大鼠视网膜外植体再生的能力就证明了这一点。
To facilitate the development of autologous transplantation techniques with which to test the ability of Schwann cell (ScC) implantations to treat nervous system injury, we have developed a method for procuring large, essentially pure populations of ScCs from adult peripheral nerve. By allowing small explants of peripheral nerve trunk to undergo axonal and myelin breakdown in vitro, rather than dissociating the nerve immediately after harvest, we are able to (1) rid the explant of nearly all fibroblasts and (2) capitalize on the intrinsic ScC mitogenic response to peripheral nerve degeneration. Here, we describe a method that yields up to 98% pure ScC populations from adult rat sciatic nerve (based on cell soma and nuclear morphology, S100 staining, and behavior of dissociated cells on neurites) at cell yields of greater than 2 x 10(4) cells/mg of starting nerve weight. The purification technique was successfully applied to human tissue; human phrenic nerve yielded 98% pure ScC populations at cell yields of 2 x 10(4) cells/mg of initial nerve weight. Similar to neonatally derived ScCs, adult rat cells can be expanded in coculture with dorsal root ganglion (DRG) neurons or in isolation in the presence of glial growth factor and forskolin. Cells expanded indefinitely on DRG neurons, or up to 10 weeks on chemical mitogens, return to quiescence following removal of the mitogenic stimulus. Expanded adult-derived rat ScCs retain functional capacity, as evidenced by their ability to myelinate DRG neurites and to support regeneration of processes from embryonic rat retinal explants.