Department of Animal Sciences, Purdue University, West Lafayette, Indiana, 47907, USA.
Cytotechnology. 1997 Jan;23(1-3):221-30. doi: 10.1023/A:1007915618413.
The zebrafish is a polular nonmammalian model for studies of neural development. We have derived cell cultures, initiated from blastula-stage zebrafish embryos, that differentiate in vitro into neurons and astrocytes. Cultures were initiated in basal nutrient medium supplemented with bovine insulin, trout serum, trout embryo extract and fetal bovine serum. After two weeks in culture the cells exhibited extensive neurite outgrowth and possessed elevated levels of acetylcholinesterase enzyme activity. Ultrastructural analysis revealed that the neurites possessed microtubules, synaptic vessicles and areas exhibiting growth cone morphology. The cultures expressed proteins recognized by antibodies to the neuronal and astrocyte-specific markers, neurofilament and glial fibrillary acidic protein (GFAP). Poly-D-lysine substrate stimulated neurite outgrowth in the cultures and inhibited the growth of nonneuronal cells. Medium conditioned by the buffalo rat liver line, BRL, promoted the growth and survival of the cells in culture. Mitotically active cells were identified in cultures that had undergone extensive differentiation. The embryo cell cultures provide an in vitro system for investigations of biochemical parameters influencing zebrafish neuronal cell growth and differentiation.
斑马鱼是一种常用于神经发育研究的非哺乳动物模式生物。我们从斑马鱼囊胚期胚胎中分离出的细胞培养物在体外分化为神经元和星形胶质细胞。培养物在基础营养培养基中起始培养，添加牛胰岛素、鳟鱼血清、鳟鱼胚胎提取物和胎牛血清。培养两周后，细胞表现出广泛的神经突生长，并具有较高水平的乙酰胆碱酯酶活性。超微结构分析显示，神经突具有微管、突触小泡和表现出生长锥形态的区域。培养物表达了神经元和星形胶质细胞特异性标志物抗体识别的蛋白质，神经丝和胶质纤维酸性蛋白（GFAP）。多聚-D-赖氨酸底物刺激培养物中的神经突生长，并抑制非神经元细胞的生长。BRL 布法罗大鼠肝系条件培养基促进细胞在培养中的生长和存活。在经历广泛分化的培养物中鉴定出有丝分裂活性细胞。胚胎细胞培养物为研究影响斑马鱼神经元细胞生长和分化的生化参数提供了体外系统。
