The effect of CHO cell initial seeding density on the results of cytotoxicity test was investigated. The doubling time of CHO cells with different initial seeding densities were determined. Neutral red assay and MTT assay were used to test the cytotoxicity of different density cells. Results showed, with 2 × 104 -6 × 104 CFU · mL-1 as the initial seeding density,the growth rate of CHO cell was relatively stable within 48 h,with 4 × 104-6×104 CFU · mL-1 as the initial seeding density, neutral red toxicity test results was more reliable, with 2×104-4×104 CFU · mL-1 as the initial seeding density,MTT toxicity test results was more reliable. The proper cell initial seeding density was beneficial to improving the reliability and repeatability of cytotoxicity test results.%考察了CHO细胞初始接种密度对体外细胞毒性试验结果的影响.通过细胞倍增试验检测不同初始接种密度下CHO细胞倍增时间的差异,并分别采用中性红比色法及MTT比色法检测CHO细胞初始接种密度对细胞毒性试验结果的影响.结果表明,CHO细胞初始接种密度对细胞毒性试验的结果存在一定影响:以2×104～6×104个·mL-1为细胞初始接种密度,CHO细胞在48 h内生长速度较为稳定；以4×104～6×104个·mL-1为细胞初始接种密度,中性红比色法毒性试验结果较为稳定；以2×104～4×104个·mL-1为细胞初始接种密度,MTT比色法细胞毒性试验结果较为稳定.在不同的细胞毒性试验中选择适合的细胞初始接种密度有助于提高细胞毒性试验结果的可重复性.
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