脐带间充质干细胞体内示踪技术的建立及在BALB/c裸鼠体内生物分布研究  

Establishment of the method for stem cells labeling and tracking of umbilical cord-mesenchymal stem cells in BALB/c nude mice

作　　者：黄瑛[1] 张澄[1] 秦超 霍艳[1] 王三龙[1] 耿兴超[1] HUANG Ying;ZHANG Cheng;QIN Chao;HUO Yan;WANG San-long;GENG Xing-chao(National Center for Safety Evaluation of Drugs,National Institutes for Food and Drug Control,The Beijing Key Lab for Pre-Clinical Safety Evaluation of Drugs,Beijing 100176,China)

机构地区：[1]中国食品药品检定研究院国家药物安全评价监测中心/药物非临床安全评价研究北京市重点实验室,北京100176

出　　处：《中国医药生物技术》2024年第6期511-520,共10页Chinese Medicinal Biotechnology

基　　金：国家重点研发计划(2021YFA1101602);中国食品药品检定研究院关键技术研究基金(GJJS-2022-6-1)。

摘　　要：目的建立适用于脐带间充质干细胞(UC-MSC)标记和体内示踪技术,并考察UC-MSC在BALB/c裸鼠体内分布特征。方法采用DiR染料标记脐带间充质干细胞,系统评价荧光染料DiR标记对UC-MSC活率、形态、表面标志物、细胞周期等影响以及进行动物体内示踪的可行性。在此基础上将标记的UC-MSC尾静脉注射BALB/c裸鼠,剂量为3×10^(6)个/只,利用活体成像技术、免疫荧光等方法检测UC-MSC在BALB/c裸鼠体内的分布代谢情况。结果DiR在5μg/ml浓度下标记5 min,干细胞标记率高,且对细胞形态、细胞活性、表面标志物、细胞周期均无明显影响;尾静脉注射后UC-MSC迅速分布在肺脏和肝脏,随后在脾脏等脏器分布。移植后30 d活体成像和免疫荧光方法均已检测不出UC-MSC。结论DiR可成功标记UC-MSC并对其多种生物学指标无明显影响;UC-MSC回输小鼠后,在体内不会广泛分布和长期存续,具有较好的临床前安全性。Objective To establish the method for labeling and tracing umbilical cord mesenchymal stem cells(UC-MSCs)in vivo and to investigate the distribution of UC-MSCs in BALB/c nude mice.Methods DiR stain was used to label the UC-MSCs.The effects of DiR labeling on the viability,morphology,surface markers and cell cycle of UC-MSCs were evaluated.On this basis,the DiR labeled UC-MSCs were injected into BALB/c nude mice by tail vein at a dose of 3×10^(6) cells.The distribution of UC-MSCs in BALB/c nude mice were detected by in vivo imaging and immunofluorescence.Results The optimal concentration and labeling time for DiR was 5μg/ml and 5 min,respectively.There were no significant changes in cell morphology,cell activity,surface markers and cell cycle.After injection,UC-MSCs were rapidly distributed in the lung and liver,and subsequently in the spleen and other organs.UC-MSCs could not be detected by the living imaging and immunofluorescence at 30 days after transplantation.Conclusion DiR stain can be used to label UC-MSCs and has no significant effect on the cell biological characteristics of UC-MSCs.UC-MSCs cannot be widely distributed and remain for a long time in vivo after transplantation in mice,suggesting that UC-MSCs has a good preclinical safety.

关 键 词：间充质干细胞 体内示踪技术 成像 生物分布 非临床研究 

分 类 号：R318[医药卫生—生物医学工程]