摘要
设计采用NAG酶活性荧光测定法检测3T3细胞增殖,本实验确立了检测3T3细胞NAG酶活性的一些实验条件,并将该方法用于bFGF的生物活性鉴定。结果:(1)3T3细胞数目在10~3-1.5×10~5之间与NAG酶活性成直线关系,且检测下限细胞数目低达10~3个细胞,表明该方法灵敏度高。(2)接种3T3细胞密度以10~3-1.5×10~3/孔(96孔培养板)为宜。(3)维持3T3细胞正常成活的最适血清浓度为0.4％—0.8％。(4)检测3T3细胞NAG酶活性以样品刺激培养48h为宜。(5)bFGF样品与标准品一样能促进3T3细胞增殖,但bFGF样品生物活性较bFGF标准品差。结论:NAG酶活性测定法能用来检测3T3细胞增殖,该方 法优于以往的细胞计数法、~3H—TdR参入法。
In the present study 3T3 cell numbers were determined by monitaring its N-acetyl-β-D-glucosaminidase activity. We studied the experimental condition of the method,and the bioactivity of bFGF was determined by the method. It was found that 3T3 cell numbers are proportional to its N-acetyl-β-D-glucosaminidase activity. The experimental condition:1. 3T3 cells were seeded to 96-well Micro Test plate about 103-1. 5×103 per well. 2. 3T3 cells has a maximal response to bFGF when they were maintained in a low serum concentration between 0. 4% and 0.8% calf serum. 3. N-acetyl-β-D-glucosaminidase activity of 3T3 cells should be monitored after 48 hours of culture. 4. bFGF samples stimulated 3T3 cells proliferation as bFGF Standard. Therefore 3T3 cell numbers can be determined by monitoring its N-acetyl-β-D-glucosaminidase activity.
出处
《细胞生物学杂志》
CSCD
1998年第1期41-45,共5页
Chinese Journal of Cell Biology
基金
全军95青年基金部分资助
课题编号96Q017
关键词
NAG
酶活性
3T3细胞
细胞增殖
荧光测定法
4-methyl-umbelliferyl-N-acetyl-β-D-glucosaminide 3T3 cell Basic fibroblast growth factor
