摘要
目的 在培养的心脏成纤维细胞模型上建立检测增殖的方法。 方法 (1)差速贴壁分离法培养乳鼠心脏成纤维细胞 ,用光镜、电镜及免疫细胞化学染色进行鉴定。 (2 )同步测定心脏成纤维细胞在不同浓度胎牛血清作用下 Brd U掺入和 WST- 1代谢活性。 结果 (1)培养的细胞经形态学观察和波形蛋白 SP染色 ,鉴定为成纤维细胞。 (2 )在 0 ,1% ,2 .5 % ,5 %和 10 %的胎牛血清作用下 ,心脏成纤维细胞 Brd U ,WST- 1的 OD值分别是 0 .186± 0 .0 2 5 ,0 .30 0± 0 .0 73;0 .46 6± 0 .0 49,0 .5 31± 0 .0 72 ;0 .76 1± 0 .0 39,0 .716± 0 .0 6 4;0 .834± 0 .0 48,0 .833±0 .0 73和 0 .90 5± 0 .0 6 4,0 .935± 0 .0 5 3(P<0 .0 1)。 结论 (1) WST- 1可在活的心脏成纤维细胞代谢 ,WST- 1法适用于该细胞的增殖。 (2 ) Brd U、WST- 1EL ISA可同步测定心脏成纤维细胞的 DNA合成及其代谢活性 ,是检测细胞增殖的新方法。
Objective To establish a new procedure for the quantification of cell proliferation on the cultured cardiac fibroblasts. Methods (1)The neonatal rat cardiac fibroblasts were cultured by the differential attachment technique and were identified with light microscopy,electromicroscopy and immunocytochemistry. (2)At different concentrations of fetal calf serum(FCS),BrdU incorporation and WST\|1 metabolic activity on the cultured neonatal rat cardiac fibroblasts were measured simultaneously with BrdU,WST\|1 ELISA. Results (1)The cultured cells identified by morphology and SP staining with vimentin showed the characteristic of fibroblasts. (2)At 0, 1%, 2 5%, 5% and 10% FCS,the absorbance spectra(optical density) of BrdU incorporation and WST\|1 cleavage products on the cultured cardiac fibroblasts were following respectively:0 186±0 025, 0 300±0 073; 0 466±0 049, 0 531±0 072; 0 76±0 039, 0 716±0 064; 0 834±0 048, 0 833±0 073 and 0 905±0 064, 0 935±0 053(P<0 01). Conclusion (1)WST\|1 can be metabolized by the viable cardiac fibroblasts,thus WST\|1 ELISA is suitable for proliferation research of this cell type. (2)The combination of BrdU and WST\|1 ELISA can be used for the simultaneous measurment of DNA synthesis and metabolic activity and as a new procedure for the quantification of cell proliferation of cardiac fibroblasts.\;
出处
《福建医科大学学报》
2000年第1期10-13,共4页
Journal of Fujian Medical University
基金
福建省科委科技基金!99-Z-163
关键词
心脏
成纤维细胞增殖
体外培养
DNA合成
myocardium cytology
cytodiaeresis
DNA synthesis
fibroblast
cell cycle
