保存温度和时间对外周血样本染色体制备的影响

Impact of peripheral blood sample storage temperature and duration on the chromosomal preparation

目的 探索外周血样本保存温度和保存时间对染色体G显带核型制备效果的影响。方法 以有核细胞数、有丝分裂指数和分裂相细胞数作为评价指标,分别对4℃和室温(RT,18~22℃)条件下,保存不同时间(0、3、5、7 d)的外周血样本染色体制备效果进行评估。结果 4℃或RT条件下,收获的有核细胞数量均随着样本保存时间的延长而减少。与新鲜外周血样本相比,4℃保存7 d以内或RT保存3 d以内的样本,有丝分裂指数分别为6.2 (1.8)%vs 7.2(1.9)%、8.2(2.0)%vs 7.2(1.9)%;4℃条件下保存3 d的样本,有丝分裂指数为9.7(2.9)%vs 7.2(1.9)%,以上差异均没有统计学意义(P>0.05)。4℃保存7 d以内或RT保存3 d以内的样本,均可获得足够数量的分裂相细胞用于染色体核型分析,其中4℃保存3 d以内的样本收获的分裂相细胞与新鲜样本相比,差异没有统计学意义[10.7(3.2)×104vs 11.4(3.5)×104,P>0.05]。结论 4℃保存7 d以内或RT保存3 d以内的外周血样本,均可以进行染色体核型制备。

Objective To explore the impact of peripheral blood sample storage temperature and duration on the quality of chromosome G banding karyotype preparation. Methods The indexes of number of nucleated cells, mitotic index and number of mitotic phase cells were used to assess the quality of preparation with the samples stored at 4 ℃ and RT(room temperature, 18-22℃) for various time durations(0, 3, 5, 7 days), respectively. Results The number of nucleated cells harvested decreased with increasing storage duration at 4 °C or RT conditions. The mitotic index did not decrease significantly for samples stored at 4 ℃within 7 days and at RT within 3 days [6.2(1.8)% vs 7.2(1.9)%, 8.2(2.0)% vs 7.2(1.9)%, both P>0.05];the samples stored at 4 ℃for 3 days showed a slight increase in the mitotic index, but the difference was not significant [9.7(2.9)% vs 7.2(1.9)%, P>0.05].Storing samples at 4 ℃ within 7 days or at RT within 3 days were still abel to provide sufficient mitotic phase cells for karyotype analysis. Additionally, the difference in mitotic phase cells between samples stored at 4 ℃ for up to 3 days and fresh samples was not statistically significant [10.7(3.2)×10~4vs 11.4(3.5)×10~4, P>0.05]. Conclusions Peripheral blood samples stored at 4 ℃within 7 days or at RT within 3 days can be used for chromosome preparation.