流式细胞术测量活性氧以评估全身预处理照射对 NOG 小鼠的影响。
Flow Cytometric Measurement of Reactive Oxygen Species to Assess the Effects of Preconditioning Total Body Irradiation on NOG Mice.
机构信息
Department of Applied Cell Sciences, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran.
Gene Therapy Research Center, Digestive Disease Research Institute, Tehran University of Medical Sciences, Tehran, Iran.
出版信息
Asian Pac J Cancer Prev. 2022 Feb 1;23(2):383-388. doi: 10.31557/APJCP.2022.23.2.383.
BACKGROUND
Preclinical development of new drugs for cancer immunotherapy requires preconditioning total body irradiation (TBI) of mice to be humanized via hematopoietic stem cell transplantation. To assess the effect of preconditioning TBI, we detected the reactive oxygen species (ROS), Annexin V, propidium iodide (PI) level in bone marrow samples by flow cytometer.
METHODS
We divided all NOG mice between irradiated (n = 20) and control groups (n = 10) for two time points. Irradiated mice were exposed to 3.5 Gy of radiation. After sacrificing BM samples were collected, the flow cytometric percentage of ROS, Annexin V, and PI markers were investigated on days 2 and 14 after exposure.
RESULTS
At the first time point, the level of ROS was higher in the irradiated group than in the control group, and this difference was statistically significant (P < 0.05). Also, at the second time point, the mean differences of all markers in the irradiated group were significantly compared to the control group (P < 0.05).
CONCLUSION
Thus, in NOG mice, the measurement of ROS level is helpful to the assessment of preconditioning TBI.
背景
癌症免疫疗法新药的临床前开发需要通过造血干细胞移植使小鼠全身照射（TBI）人源化。为了评估预处理 TBI 的效果，我们通过流式细胞仪检测骨髓样本中的活性氧（ROS）、Annexin V、碘化丙啶（PI）水平。
方法
我们将所有 NOG 小鼠分为照射组（n = 20）和对照组（n = 10），共两个时间点。照射组小鼠接受 3.5 Gy 的辐射。在牺牲后采集 BM 样本，在暴露后第 2 天和第 14 天通过流式细胞术检测 ROS、Annexin V 和 PI 标志物的百分比。
结果
在第一个时间点，照射组的 ROS 水平高于对照组，差异具有统计学意义（P < 0.05）。此外，在第二个时间点，照射组的所有标志物的平均差异与对照组相比均具有显著差异（P < 0.05）。
结论
因此，在 NOG 小鼠中，ROS 水平的测量有助于评估预处理 TBI。
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