Development and characterization of cell lines from heart, liver, spleen and head kidney of sea perch Lateolabrax japonicus
Development and characterization of cell lines from heart, liver, spleen and head kidney of sea perch Lateolabrax japonicus
复制标题
DOI：
10.1111/j.1095-8649.2006.01155.x
复制
发表时间：
2006-09
影响因子：
2
通讯作者：
Hanqing Ye;Songlin Chen;Z. Sha;Meiyu Xu
中科院分区：
文献类型：
--
作者：
Hanqing Ye;Songlin Chen;Z. Sha;Meiyu Xu
以日本鲈鱼（Lateolabrax japonicus）头肾、脾、肝和心为材料，建立了LJHK、LJS、LJL、LJH-1和LJH-2 5个细胞系。将LJHK、LJS、LJL、LJH-1和LJH-2细胞系在添加胎牛血清、海鲈血清和10 ng ml -1碱性成纤维细胞生长因子（bFGF）的最低基本培养基（MEM）中传代46、32、32、36和34次。用显微镜连续观察5种细胞系的原代培养和传代培养的形态。所有细胞系的生长适宜温度均为18 ~ 30℃，24℃时生长最佳，<18℃时生长速率降低，FBS的最佳浓度为10%，培养基中添加bFGF可显著提高细胞的生长速率。在24℃条件下，在1 × 105 cells ml -1的镀密度下，LJS、LJH-1、LJL、LJH-2和LJHK细胞的倍增时间分别为52-7、54·9、57、58-7和66 h。染色体分析显示，LJH-1、LJH-2、LJHK、LJL和LJS细胞系中分别有42、48、38、43和45%的细胞维持正常的二倍体染色体数（48）。用绿色荧光蛋白（GFP）报告质粒转染LJHK细胞成功，细胞中绿色荧光蛋白基因的表达表明了该细胞在基因表达研究中的潜在应用价值。此外，脂多糖处理LJHK细胞可导致IL-1β的表达增加，这表明LJHK细胞可能是研究鱼类免疫调节基因表达和功能的有价值的工具。
Five cell lines (LJHK, LJS, LJL, LJH-1 and LJH-2) were established from the head kidney, spleen, liver and heart of sea perch Lateolabrax japonicus. The cell lines LJHK, LJS, LJL, LJH-1 and LJH-2 were subcultured 46, 32, 32, 36 and 34 times in minimum essential medium (MEM) supplemented with foetal bovine serum (FBS), sea perch serum and 10 ng ml -1 basic fibroblast growth factor (bFGF). Morphology of primary cultures and subcultures of the five cell lines were observed continuously by microscopy. The suitable temperature for growth was 18 to 30° C for all of these cell lines with the optimum growth at 24° C and a reduced growth rate <18° C. The optimum concentration of FBS was found to be 10% and addition of bFGF to the medium significantly increased the growth rate of the cells. The doubling time of LJS, LJH-1, LJL, LJH-2 and LJHK cells was determined to be 52-7, 54·9, 57, 58-7 and 66 h at a plating density of 1 x 10 5 cells ml -1 at 24° C, respectively. Chromosome analysis revealed that 42, 48, 38, 43 and 45% cells maintained normal diploid chromosome number (48) in the LJH-1, LJH-2, LJHK, LJL and LJS cell lines, respectively. The LJHK cells were successfully transfected with green fluorescent protein (GFP) reporter plasmids and the expression of GFP gene in the cells indicated the possible utility of the cells in gene expression studies. Furthermore, treatment of the LJHK cells with lipopolysaccharide led to increased expression of IL-1β, demonstrating that LJHK cells might be a valuable tool for studying the expression and function of immunomodulatory gene in fishes.