Woodhams P L, Wilkin G P, Balàzs R
Dev Neurosci. 1981;4(4):307-21. doi: 10.1159/000112770.
Astrocyte-enriched monolayer cultures were obtained by growing cells dissociated from 8-day-old rat cerebella pretreated with hydroxyurea. After 7 days in vitro a carpet of flat polygonal cells was obtained. The identity of these flat cells was established and their numbers quantified using immunological markers for astrocytes, fibroblasts and endothelial cells (antisera against fibrillary acidic protein and Thy-1, and Chagasic sera respectively). A combination of (3H] thymidine autoradiography and immunocytochemistry showed extensive replication of astrocytes, the number of which rose from about 20 to 80% of the total cells during the 1st week in vitro: 80% of these cells were labelled at 7 days after exposure to thymidine on the 1st day in culture. The culture conditions chosen did not support the survival of nerve cells. and fibroblasts constituted 5-7% of the cells at 7-14 DIV, whilst the total cell number remained roughly constant. On the other hand, staining with a new specific marker for endothelial cells revealed that these cells divided actively, ultimately accounting for the majority of the non-astrocytic cells.
通过培养从经羟基脲预处理的8日龄大鼠小脑分离的细胞，获得了富含星形胶质细胞的单层培养物。体外培养7天后，得到了一层扁平的多边形细胞。利用针对星形胶质细胞、成纤维细胞和内皮细胞的免疫标记物（分别为抗纤维酸性蛋白和Thy-1的抗血清以及恰加斯血清）确定了这些扁平细胞的身份并对其数量进行了量化。[3H]胸腺嘧啶核苷放射自显影术和免疫细胞化学相结合的方法显示星形胶质细胞大量增殖，在体外培养的第1周内，其数量从约占总细胞数的20%增加到80%：在培养第1天接触胸腺嘧啶核苷后7天，这些细胞中有80%被标记。所选择的培养条件不支持神经细胞的存活。在培养7 - 14天（DIV）时，成纤维细胞占细胞总数的5 - 7%，而细胞总数大致保持不变。另一方面，用一种新的内皮细胞特异性标记物染色显示，这些细胞活跃分裂，最终占非星形胶质细胞的大多数。
