黄鳍鲷肝细胞体外培养研究

Primary Culture of Liver Cell from Sparus latus hottuyn in vitro

以黄鳍鲷肝脏细胞为材料 ,研究其肝细胞体外培养适宜的 p H、血清浓度、谷氨酰胺等条件 ,并进行传代培养试验。结果表明 ,培养液为 :MEM含 2 0 %小牛血清、15 g/ L L-谷氨酰胺及 p H 6 .9～ 7.1时 ,在 2 3℃中静置培养 ,黄鳍鲷肝细胞生长良好 ,3～ 5 d长满单层 ,细胞主要为成纤维细胞状 ,并能顺利传代。细胞动力学试验表明 :传代细胞 1～ 2代为适应期 ,3～ 5代为旺盛期 。

Primary cultures and subculture of liver cells from Sparus latus houttuyn in vitro were studied. MEM medium with 20% calf serum and L Glutamine 15 g/L,pH 6.9 7.1were found to provide the best growing conditions of cells. The culture temperature was 23 ℃. Under the this conditions, monolayers of primarily cultured liver cells were routinely obtained with 100% confluence within 3 to 5 days, and the primarily cultured cells can also be successful subculture under the same condition. The cultured cell morphology was mainly fibroblast like. The cell kinetic assay indicates that there was an adaptable growth period in 1 st 2 nd generations, a fast growth period in 3 rd 5 th generations and a degenerating period behind 6 th generations.