目的 探索小鼠淋巴瘤胸腺激酶（tk）位點基因突變試驗中血清滅活對結(jié)果的影響。方法 取對數(shù)生長期的小鼠淋巴瘤細(xì)胞（L5178Y3.7.2c-tk^+/-）設(shè)立陰性對照組和陽性對照組，陰性對照組加入100 μL DMSO，陽性對照組加入100 μL 10 μg/mL 4-硝基喹啉（4-NQO）誘導(dǎo)突變，測定細(xì)胞懸浮增長率（SG）、相對懸浮增長率（RSG）、平板效率（PE）、相對存活率（RS）和相對總生長率（RTG）等指標(biāo)。根據(jù)血清滅活與否和選擇突變劑三氟胸苷（TFT）的含量，在篩選突變集落過程中，將陰性對照組和陽性對照組各分為：滅活組、未滅活+1.5倍TFT組、未滅活組，測定突變頻率（MF），對試驗結(jié)果進(jìn)行評價。結(jié)果 陰性對照組SG在8~32，細(xì)胞倍增速度符合試驗要求；PE在65%~120%，符合試驗成立條件。陽性對照的滅活組、未滅活+1.5倍TFT組、未滅活組中MF均比陰性對照組至少增加300×10^-6且小克隆突變頻率至少占40%，符合試驗要求。陰性對照滅活組的MF數(shù)值在標(biāo)準(zhǔn)范圍內(nèi)，未滅活組的MF數(shù)值明顯高于滅活組，超過標(biāo)準(zhǔn)數(shù)值范圍，且96孔板中的大小克隆不易區(qū)分；陰性對照未滅活+1.5倍TFT組使得MF數(shù)值在標(biāo)準(zhǔn)范圍內(nèi)，但遠(yuǎn)高于滅活組。結(jié)論 在小鼠淋巴瘤試驗中，測定MF的平板一定要用滅活的血清，未滅活血清可能通過抵消TFT作用對結(jié)果產(chǎn)生很大影響。

Objective To explore the effect of inactivated serum on the mouse lymphoma tk gene mutation assay. Methods The negative control group and positive control group were set up for the mouse lymphoma cells (L5178Y3.7.2c-tk^+/-) during the logarithmic growth period. The negative control group was added to 100 L DMSO. The positive control group added 100 μL 10 μg/mL 4-nitroquinoline (4-NQO) induced mutation, and the cell suspension growth rate (SG), the relative suspension growth rate (RSG), plate efficiency (PE), relative survival rate (RS), relative total growth rate (RTG), and other indicators were measured. According to the inactivation of serum and the content of three fluorothoracoside (TFT), the negative control group and the positive control group were divided into inactivation group, not inactivated + 1.5×TFT group and the not inactivated group. We measured the mutation frequency (MF) value and evaluated the test results. Results In negative control group, SG was from 8 to 32, and cell doubling speed accords with test requirement. PE from 65% to 120% conforms to the established condition of experiment. MF in positive control group increased by 300×10^-6 than the negative control group, and the frequency of the small clone mutation was at least 40%, which was in line with the test requirements. MF value of the negative control inactivation group was within the standard range. The MF value of the not inactivated group was significantly higher than that of the inactivated group, exceeding the standard number, and Banzhong size clones were not easy to distinguish; The negative control not inactivated +1.5×TFT group made the MF value within the standard range, but much higher than the inactivated group. Conclusions In mouse lymphoma test, the plates with measuring mutation frequency must use the heat inactivate serum, and the not inactivated serum may have a great effect on the results by counteracting TFT.

國家科技重大新藥創(chuàng)制項目（2015ZX09501004）；天津市科技計劃項目（16PTGCCX00090）