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gary286

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[求助] 审稿人问细胞毒性。why the sample has less viability at 4h than 8h,24h?

审稿人问细胞毒性。why the sample has less viability at 4h than 8h,24h?

无机材料用MTT法做细胞毒性,4小时的cell viability 明显小于8小时和24小时的。审稿人问原因,审稿人很可能也不是搞生物的。我能简单回答是误差吗(experiment error)?本人对细胞毒性检测不太了解,见笑了,希望大家帮忙看下,如何给出完备的回答呢。。。

PS, 无机材料为生物相容性材料修饰的Fe3O4。

实验方法如下,
The cytotoxicity study was measured using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in MCF-7 cells. MCF-7 cells were maintained in DMEM media supplemented with 10% fetal bovine serum (FBS) (Gibco, Grand Island, NY., USA), 2 mM L-glutamine, 100 mg/ml streptomycin, and 100 U/ml penicillin. The cell lines were cultured in 35 mm cover glass-bottom dish at 37 oC in a humidified atmosphere containing 5% CO2. The magnetite particles at concentrations of 0, 0.03, 0.3 mg/mL in DMEM medium were mixed with the cells, and the cells were incubated for 4 h, 8 h, 24 h, 48 h, and 96 h. Optical density of each sample was measured in a Dynatech MR7000 ELISA reader at 570 nm. The optical density of the media was proportional to the number of viable cells. Inhibition of proliferation was evaluated as a percentage of control growth. The results were expressed as an average over three nominally identical measurements.

结果如下:
0.03    4~96h           97.33263;  96.01693;96.71721;97.61354;95.56296%
0.3      4~96h         89.94877; 93.27308;94.00933;94.81344;91.00256 %
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gary286(金币+18): 多谢斑竹,MTT的重复性 可作为突破口。。MTT 优点应该是快速,方便吧? 2011-06-13 19:33:17
gary286(金币+10): 2011-06-30 22:55:47
怀疑误差,你做了几复孔,几平行,几重复?
MTT本来重复性就极差
2楼2011-06-13 18:48:38
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