非洲爪蟾肝细胞原代培养方法

非洲爪蟾是研究内分泌干扰物的良好模型动物,其体外肝细胞可用于类雌激素活性评价、污染物代谢等研究.论文探讨了非洲爪蟾肝细胞原代培养的方法,采用两步原位灌注法分离非洲爪蟾肝细胞,通过胶原酶的作用使细胞之间解离,后经过一系列转速的离心,获得纯化的肝实质细胞.研究结果表明,采用此法获得的细胞数量为2.5～5×106个,细胞成活率达95%以上,纯度在95%以上,细胞胞体透亮,折光性强,状态良好.培养24h后贴壁较好,每2d换液1次,可培养8～10d,细胞可满足多种后续实验的要求.

Primary Cultured Protocol on Hepatocytes of Xenopus laevis

Xenopus laevis is a good model to study endocrine disrupters, and the hepatocytes are widely used for assaying estrogenic activity and studying metabolism of endocrine disruptors. The present study aims to introduce a primary cultured protocol on hepatocytes of Xenopus laevis. A two-step in situ perfusion method was used. The collagenase was used to digest collagen in the liver tissue, and the centrifugation steps followed to free the dispersed cells from unwanted tissue material and the erythrocytes. The final cell preparation was seeded at a density of 105/ml, and about 2.5~5×106 cells were obtained totally. The cells could be well-maintained for 8~10 days by replacing the culture medium every 2 days. The primary cultured hepatocytes could be well subjected to lots of follow-up experiments.