Wrathall J R, Rigamonti D D, Braford M R, Kao C C
Brain Res. 1981 Dec 14;229(1):163-81. doi: 10.1016/0006-8993(81)90753-8.
There are several methods available for the production of Schwann cell cultures from fetal and neonatal peripheral nervous tissue. We have investigated methods for producing Schwann cell-rich cultures from adult tissue. Dorsal root ganglia from normal adult cats were used to initiate explant cultures or subjected to primary dissociation. The resulting cultures were compared in terms of growth, the proportions of fibroblastic and Schwann-like cells in primary cultures and the effects of subculture on the relative frequency of these cell types. We found that excision and transfer of explanted ganglion pieces after 14 days in culture produced a secondary outgrowth rich in small, bipolar, spindle-shaped Schwann-like cells. Subculture of this outgrowth produced secondary cultures of predominantly Schwann-like cells with typical spindle-shaped morphology. The use of antimitotic agents in the media to inhibit fibroblast growth was not observed to be necessary or beneficial with this adult tissue. Primary dissociation of ganglia with enzymes (trypsin or collagenase) and mechanical agitation was even more effective in producing secondary cultures and cell lines that were, by morphological criteria, predominantly or exclusively Schwann-like cells. One of these Schwann-like cell lines, designated GSA, has been carried over 24 subcultures while retaining characteristic Schwann cell morphology. Cells of this line have been examined by scanning and transmission electron microscopy. Karyotype analysis indicates a chromosome complement consistent with the species of origin, a normal cat.
有几种方法可用于从胎儿和新生儿外周神经组织中培养雪旺细胞。我们研究了从成年组织中培养富含雪旺细胞的培养物的方法。使用正常成年猫的背根神经节来启动外植体培养或进行原代解离。对所得培养物在生长、原代培养中成纤维细胞和雪旺样细胞的比例以及传代培养对这些细胞类型相对频率的影响方面进行了比较。我们发现,培养14天后切除并转移外植的神经节片段会产生富含小的、双极的、纺锤形雪旺样细胞的二次生长。这种二次生长的传代培养产生了主要为具有典型纺锤形形态的雪旺样细胞的二次培养物。在这种成年组织中,未观察到在培养基中使用抗有丝分裂剂来抑制成纤维细胞生长是必要的或有益的。用酶(胰蛋白酶或胶原酶)和机械搅拌对神经节进行原代解离,在产生二次培养物和细胞系方面甚至更有效,根据形态学标准,这些二次培养物和细胞系主要或完全是雪旺样细胞。其中一个雪旺样细胞系,命名为GSA,已经传代24次,同时保留了雪旺细胞的特征形态。已通过扫描电子显微镜和透射电子显微镜对该细胞系的细胞进行了检查。核型分析表明染色体组成与来源物种一致,即正常的猫。