JMP定制实验设计优化胃癌患者PBMC分离方案

The Optimization of PBMC Isolation Condition in Gastric Cancer Patients Based on Custom Designs of JMP Software

为探索胃癌患者外周血单个核细胞(peripheral blood mononuclear cell,PBMC)分离的优化条件,本研究用JMP10.0软件定制设计方法,研究离心时间、离心速度对PBMC离心厚度的影响,建立模型、刻画最优参数,验证并用台盼蓝染色观察优化条件下离心的PBMC细胞存活率。结果表明,离心时间、离心速度与PBMC厚度存在曲线关系,离心时间与离心速度的交互作用无统计学意义。预测模型的决定系数R2=0.88,预测值与实际值的一致性较好。刻画最优离心时间=10 min,离心速度=2 307 r/min,PBMC厚度预测值=2.596 923 mm,95%CI(2.342 13,2.851 71),验证离心的PBMC厚度=2.7 mm,细胞存活率=95%。本研究采用JMP定制设计方法优化PBMC离心条件,可为标准操作程序的建立提供基础。

In order to explore the optimal conditions for isolation of peripheral blood mononuclear cells （PBMC） from gastric cancer patients＇ peripheral blood, custom designs of JMP software （10.0） were used in this study to ex- plore the effect of centrifugal time and velocity on PBMC thickness. Besides, the model was optimized, the cen- trifugal parameters were established, and the survival rate of acentric PBMC cells under the optimized conditions was observed by trypan blue staining. The results showed that there was a curvilinear relationship among centrifu- gal time, velocity and the thickness of PBMC, and the interaction between centrifugal time and velocity was not statistically significant. The decision coefficient of the prediction model was W=0.88, which indicated that the pre- dicted values were in good agreement with the actual values. The predicted optimal centrifugal time was l 0 min, centrifugal velocity was 2 308 r/min, predicted PBMC thickness was 2.596 911 mm, 95% CI （2.342 15, 2.851 6＂7）, the centrifugal PBMC thickness got from verifying isolation test was 2.7 ram, and the cell survival rate was 95%. In conclusion, this study adopted JMP custom design method to optimize PBMC centrifugal conditions, which could provide basis for the establishment of standard operation procedure （SOP） for PBMC isolation.