Collodi P, Kamei Y, Ernst T, Miranda C, Buhler D R, Barnes D W
Department of Biochemistry and Biophysics, Oregon State University, Corvallis 97331.
Cell Biol Toxicol. 1992 Jan-Mar;8(1):43-61. doi: 10.1007/BF00119294.
The zebrafish is a popular model for studies of vertebrate development and toxicology. However, in vitro approaches with this organism have not been fully exploited because cell culture systems have been unavailable. We developed methods for the culture of cells from blastula-stage diploid and haploid zebrafish embryos, as well as cells from the caudal and pelvic fin, gill, liver, and viscera of adult fish. The haploid embryo-derived cells differentiated in culture to a pigmented phenotype and expressed, upon exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin, a protein that was immunologically and functionally similar to rainbow trout cytochrome P450IA1. Zebrafish cultures were grown in a complex basal nutrient medium supplemented with insulin, trout embryo extract, and low concentrations of trout and fetal bovine serum; they could not be maintained in conventional culture medium containing a high concentration of mammalian serum. Using calcium phosphate-mediated transfection, a plasmid constructed for use in mammalian cells was introduced into zebrafish embryo cell cultures and expressed in a stable manner. These results indicated that the transfection procedures utilized in mammalian systems can also be applied to zebrafish cell cultures, providing a means for in vitro alteration of the genotype and phenotype of the cells.
斑马鱼是研究脊椎动物发育和毒理学的常用模型。然而，由于细胞培养系统不可用，这种生物体的体外研究方法尚未得到充分利用。我们开发了从囊胚期二倍体和单倍体斑马鱼胚胎以及成年鱼的尾鳍、臀鳍、鳃、肝脏和内脏中培养细胞的方法。单倍体胚胎来源的细胞在培养中分化为色素沉着表型，在暴露于2,3,7,8 - 四氯二苯并 - 对 - 二恶英时，表达一种在免疫和功能上与虹鳟鱼细胞色素P450IA1相似的蛋白质。斑马鱼培养物在添加胰岛素、鳟鱼胚胎提取物以及低浓度鳟鱼和胎牛血清的复杂基础营养培养基中生长；它们无法在含有高浓度哺乳动物血清的传统培养基中维持生长。使用磷酸钙介导的转染方法，将构建用于哺乳动物细胞的质粒导入斑马鱼胚胎细胞培养物中并稳定表达。这些结果表明，哺乳动物系统中使用的转染程序也可应用于斑马鱼细胞培养，为体外改变细胞的基因型和表型提供了一种方法。
