基于生物正交点击化学修饰吲哚菁绿用于脂肪间充质干细胞活体示踪的实验研究  

In vivo tracking of adipose tissue-derived mesenchymal stem cells labeled with indocyanine green using bio-orthogonal click chemistr

作　　者：何宏星 廖乃顺 HE Hongxing;LIAO Naishun(Laboratory Animal Center,Fujian Medical University,Fuzhou 350004,China;the United Innovation of Mengchao Hepatobiliary Technology Key Laboratory of Fujian Province,Mengchao Hepatobiliary Hospital of Fujian Medical University,Fuzhou 350025)

机构地区：[1]福建医科大学实验动物中心,福州350004 [2]福建医科大学孟超肝胆医院,福建省孟超肝胆技术联合创新重点实验室,福州350025

出　　处：《中国实验动物学报》2023年第8期993-998,共6页Acta Laboratorium Animalis Scientia Sinica

基　　金：福建省自然科学基金面上项目(2020J011152)。

摘　　要：目的探究生物正交点击化学修饰可否用于脂肪间充质干细胞(ADSC)的吲哚菁绿(ICG)标记及活体示踪研究。方法收集小鼠脂肪组织进行ADSC的分离与培养,分别添加Ac4ManNAz、DBCO-ICG进行生物正交点击化学反应,评估该方法对ADSC细胞活性的影响,并利用共聚焦显微镜观察ADSC的ICG标记情况;将标记ICG的ADSC移植到急性肝损伤小鼠体内,利用近红外二区荧光成像开展ADSC的活体示踪研究,收集肝组织进行ADSC归巢性分析;最后,评估生物正交点击化学修饰对ADSC移植治疗急性肝损伤疗效的影响。结果生物正交点击化学可将ICG探针成功地修饰于ADSC细胞内,且不影响ADSC的细胞活性;标记ICG的ADSC移植到体内后主要分布、定位在肝组织内,说明ADSC具有肝向归巢性;生物正交点击化学修饰不影响ADSC改善急性肝损伤小鼠肝功能、减少肝细胞坏死的疗效。结论生物正交点击化学可用于ADSC的ICG探针标记及活体示踪研究。Objective To explore the use of bio-orthogonal click chemistry indocyanine green(ICG)labeling of adipose tissue-derived mesenchymal stem cells(ADSC)and in vivo cell tracking.Methods ADSCs were isolated and cultured,and then incubated with N-azidoacetylmannosamine-tetraacylated(Ac4ManNAz)and Dibenzocyclooctyneindocyanine green(DBCO-ICG)using bio-orthogonal click chemistry,and their cell viability was evaluated.ICG labeling was confirmed by laser confocal microscopy imaging.After ICG labeling,in vivo ADSC tracking was performed by nearinfraredⅡfluorescence imaging in acute liver injured mice.Liver tissue sections were also collected to analyze ADSC homing.Moreover,the therapeutic effects of ICG-labeled ADSCs on serum levels of alanine aminotransferase(ALT)and aspartate transaminase(AST)and on pathological changes were also evaluated.Results ICG labeling of ADSCs could be achieved by bio-orthogonal click chemistry.Notably,ADSC viability and their therapeutic effects on acute liver injury,including serum ALT and AST and hepatic morphology,were not affected by this method.Near-infraredⅡfluorescence imaging revealed the hepatic accumulation and homing of transplanted ADSCs in vivo and ex vivo.Conclusions Bioorthogonal click chemistry may provide a promising new strategy for ADSC labeling and in vivo cell tracking.

关 键 词：生物正交点击化学 吲哚菁绿 脂肪间充质干细胞 活体示踪 

分 类 号：Q95-33[生物学—动物学]