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Comparative strategies for stem cell biodistribution in a preclinical study.
Acta Pharmacologica Sinica ( IF 8.2 ) Pub Date : 2019-11-08 , DOI: 10.1038/s41401-019-0313-x
Fang Wang 1, 2 , Zhe Wang 1, 3 , Fen Wang 4 , Kelly Dong 1, 2 , Jing Zhang 1, 3 , Yun-Juan Sun 2 , Chun-Feng Liu 4, 5 , Mei-Jie Xing 2 , Xue Cheng 2 , Su Wei 2 , Jia-Wei Zheng 6 , Xiong-Fei Zhao 6 , Xiao-Ming Wang 6 , Jie Fu 1, 3 , Hai-Feng Song 1, 3
Affiliation  

Stem cell therapy represents the potential alternative effective strategy for some diseases that lack effective treatment currently. Correspondingly, it is crucial to establish high-sensitive and reliable quantification assay for tracing exogenous cell migration. In the present study, we first used both bioluminescence imaging (BLI) indirect labeling (human norepinephrine transporter-luciferase reporter system) and 89zirconium (89Zr)-hNSCs direct labeling combined with positron emission tomography/computer tomography (PET/CT) system for tracking human neural stem cells (hNSCs) migration into the brain via nasal administration in preclinical study. But the above two methods failed to give the biodistribution profile due to their low sensitivity. Considering its superior sensitivity and absolute quantitation capability, we developed and validated the droplet digital PCR (ddPCR) targeting species-specific gene in frozen and paraffin sections, slices, and whole blood with the sensitivity of 100–200 hNSCs. Accurate and high throughput quantification could be performed using ddPCR with the coefficient of variation (CVs) of lower quality control (LQC) below 30%. In combination with immunohistochemistry and ddPCR, we confirmed the migration of hNSCs into the brain via nasal administration, which supported the efficacy of hNSCs in MPTP-treated mice, an animal model of Parkinson’s disease. In conclusion, the present study is the first to report the application of ddPCR in the pharmacokinetics profile description of tracking of hNSCs in preclinical studies.



中文翻译:

临床前研究中干细胞生物分布的比较策略。

干细胞疗法代表了一些目前缺乏有效治疗的疾病的潜在替代有效策略。相应地,建立高灵敏度和可靠的定量分析方法来追踪外源性细胞迁移至关重要。在本研究中,我们首先使用了生物发光成像 (BLI) 间接标记(人去甲肾上腺素转运蛋白-荧光素酶报告系统)和89锆(89Zr)-hNSCs 直接标记结合正电子发射断层扫描/计算机断层扫描 (PET/CT) 系统,用于在临床前研究中通过鼻腔给药跟踪人类神经干细胞 (hNSCs) 迁移到大脑中。但上述两种方法由于灵敏度低,未能给出生物分布曲线。考虑到其卓越的灵敏度和绝对定量能力,我们开发并验证了针对冷冻和石蜡切片、切片和全血中物种特异性基因的液滴数字 PCR (ddPCR),灵敏度为 100-200 hNSC。使用 ddPCR 可以进行准确和高通量的定量,其中低质量控制 (LQC) 的变异系数 (CV) 低于 30%。结合免疫组化和ddPCR,我们证实了 hNSCs 通过鼻腔给药迁移到大脑中,这支持了 hNSCs 在 MPTP 治疗的小鼠(帕金森病的动物模型)中的功效。总之,本研究首次报道了 ddPCR 在临床前研究中追踪 hNSCs 的药代动力学特征描述中的应用。

更新日期:2019-11-08
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