In vivo visualization of embryonic stem cell survival, proliferation, and migration after cardiac delivery
In vivo visualization of embryonic stem cell survival, proliferation, and migration after cardiac delivery
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DOI：
10.1161/circulationaha.105.588954
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发表时间：
2006-02-21
期刊：
影响因子：
37.8
通讯作者：
Wu, JC
中科院分区：
文献类型：
--
作者：
Cao, F;Lin, S;Wu, JC
背景 - 近期研究表明干细胞疗法可促进组织再生；然而，由于传统组织学检测和成像方式的局限性，体内干细胞监测仍然存在问题。
方法与结果 - 用携带一种新型三联融合（TF）报告基因的慢病毒载体对小鼠胚胎干细胞（ES）进行稳定转导，该报告基因由萤火虫荧光素酶、单体红色荧光蛋白和截短的胸苷激酶（fluc - mrfp - ttk）组成。与未转导的对照细胞相比，报告基因或报告探针对ES细胞的活力、增殖和分化能力没有不良影响（P = 无显著差异）。之后，将1×10⁷个携带TF报告基因的ES细胞（ES - TF）注射到成年裸鼠的心肌中（n = 20）。对照动物接受未转导的ES细胞（n = 6）。在第4天，研究动物的生物发光和正电子发射断层扫描信号分别为每球面度（sr）3.7×10⁷ ± 5.8×10⁶光子·秒⁻¹·厘米⁻²和0.08 ± 0.03%注射剂量/克（与对照组相比，P < 0.05）。从第1周到期第4周，两种信号均逐渐增强，这表明ES细胞在宿主中存活和增殖。组织学分析显示形成了心内和心外畸胎瘤。最后，与用生理盐水（1 mL/kg）处理的对照动物（n = 4）相比，腹腔注射更昔洛韦（50 mg/kg）处理的动物（n = 4）未形成畸胎瘤。
结论 - 这是第一项对稳定表达荧光、生物发光和正电子发射断层扫描报告基因的ES细胞进行特性描述，并监测ES细胞存活、增殖和迁移动力学的研究。这种多功能成像平台在干细胞疗法的基础研究和临床研究中应具有广泛的应用。
Background - Recent studies have shown that stem cell therapy can promote tissue regeneration; however, monitoring stem cells in vivo remains problematic owing to limitations of conventional histological assays and imaging modalities.Methods and Results - Murine embryonic stem (ES) cells were stably transduced with a lentiviral vector carrying a novel triple-fusion (TF) reporter gene that consists of firefly luciferase, monomeric red fluorescence protein, and truncated thymidine kinase (fluc-mrfp-ttk). ES cell viability, proliferation, and differentiation ability were not adversely affected by either reporter genes or reporter probes compared with nontransduced control cells (P = NS). Afterward, 1X10(7) of ES cells carrying the TF reporter gene (ES-TF) were injected into the myocardium of adult nude rats (n=20). Control animals received nontransduced ES cells (n=6). At day 4, the bioluminescence and positron emission tomography signals in study animals were 3.7X10(7) +/- 5.8 +/- 10(6) photons . s(-1) . cm(-2) per steradian (sr) and 0.08 +/- 0.03% injected dose/ g, respectively (P < 0.05 versus control). Both signals increased progressively from week 1 to week 4, which indicated ES cell survival and proliferation in the host. Histological analysis demonstrated the formation of intracardiac and extracardiac teratomas. Finally, animals (n = 4) that were treated with intraperitoneal injection of ganciclovir (50 mg/kg) did not develop teratomas when compared with control animals (n = 4) treated with saline (1 mL/kg).Conclusion - This is the first study to characterize ES cells that stably express fluorescence, bioluminescence, and positron emission tomography reporter genes and monitor the kinetics of ES cell survival, proliferation, and migration. This versatile imaging platform should have broad applications for basic research and clinical studies on stem cell therapy.