Paim Fabilene G, Maia Leandro, Landim-Alvarenga Fernanda da Cruz, Foresti Fausto, Oliveira Claudio
Department of Morphology, Sao Paulo State University, Botucatu 18618-689, Sao Paulo, Brazil.
Department of Animal Reproduction and Radiology, Sao Paulo State University, UNESP, Botucatu 18618-681, Sao Paulo, Brazil.
Methods Protoc. 2018 Dec 10;1(4):47. doi: 10.3390/mps1040047.
Cell culture is an excellent alternative for the maintenance of cell lines and to obtain quality chromosome preparations of fishes. However, this methodology is still little employed, mainly because of the difficulty of standardization of cell cultures. In this study, we describe a methodology for the rapid acquisition of cell lineages and mitotic chromosomes for cytogenetic studies of fish species from muscle tissue cells. Our methodology is based on the use of a gelatin film, which provides better adhesion of a large number of cells and appropriate conditions for multiplication. The cells of , used as an experimental model, with fibroblast-like morphology, showed rapid cellular proliferation, resulting in a great number of cells. Chromosomal preparations of cultured cells showed the diploid number of the species, 2n = 50 chromosomes, in 80% of the cells examined, with chromosomes intact and distended. Cell populations were cryopreserved and after being recovered, these cells maintained their proliferative capacity. The development of this methodology represents an innovation for the fish cytogenetics area and it may bring a significant contribution to the conservation and study of several groups due to the difficulty of obtaining good-quality chromosome preparations.
细胞培养是维持细胞系和获得优质鱼类染色体标本的一种极佳替代方法。然而，这种方法的应用仍然很少，主要是因为细胞培养的标准化存在困难。在本研究中，我们描述了一种从肌肉组织细胞快速获取细胞谱系和有丝分裂染色体以用于鱼类细胞遗传学研究的方法。我们的方法基于使用明胶膜，它能为大量细胞提供更好的附着以及适合增殖的条件。作为实验模型的[具体物种名称未给出]细胞，具有成纤维细胞样形态，显示出快速的细胞增殖，产生了大量细胞。培养细胞的染色体标本在80%的检测细胞中显示出该物种的二倍体数目，即2n = 50条染色体，染色体完整且舒展。细胞群体被冷冻保存，复苏后这些细胞仍保持其增殖能力。这种方法的开发代表了鱼类细胞遗传学领域的一项创新，并且由于获得高质量染色体标本存在困难，它可能会对多个群体的保护和研究做出重大贡献。
