"Firefly rats" as an organ/cellular source for long-term in vivo bioluminescent imaging
"Firefly rats" as an organ/cellular source for long-term in vivo bioluminescent imaging
复制标题
DOI：
10.1097/01.tp.0000203137.06587.4a
复制
发表时间：
2006-04-27
期刊：
影响因子：
6.2
通讯作者：
Kobayashi, E
中科院分区：
文献类型：
--
作者：
Hakamata, Y;Murakami, T;Kobayashi, E
背景。涉及使用干细胞的移植研究需要适当的体内可视化系统来监测观察期内的细胞命运。体内成像的新领域正在利用荧光和发光生物技术发展起来，涉及活体动物中复杂细胞过程的实时可视化。方法。继我们最近开发近交绿色荧光蛋白 (GFP) 转基因 (Tg) 大鼠之后，我们利用萤火虫荧光素酶创建了近交 (Lewis) Tg 大鼠。通过植皮试验评估荧光素酶的免疫原性，并通过体内发光技术监测移植物的命运。结果。荧光素酶-Tg 大鼠普遍表达该标记基因。与 GFP-Tg 来源的皮肤移植（< 10 天）相比，传统的皮肤移植显然显示出野生型大鼠对荧光素酶-Tg 大鼠皮肤的长期接受性（> 100 天）。这表明对荧光素酶蛋白的细胞免疫反应比对 GFP 的低。引人注目的是，心脏和小肠器官移植以及骨髓细胞移植显示出活力和移植接受性，这表明来自荧光素酶 Tg 大鼠的细胞和器官是可移植的，并且可以在足够长的时间内追踪它们的命运。利用免疫原性较低的荧光素酶，还检查了移植的成熟肝细胞的细胞命运。移植的肝细胞增殖并在受损肝脏中选择性地进行监测，但在健康肝脏中则不然，持续 60 多天。结论。基于这些发现，我们提出，具有现代光学成像的荧光素酶-Tg 大鼠系统为更好地了解干细胞生物学和移植提供了一个新平台。
Background. Transplantation research involving the use of stem cells demands an appropriate in vivo visualization system to monitor cellular fate over an observation period. The new field of in vivo imaging is being developed with fluorescent and luminescent biotechnology, and involves the real-time Visualization of complex cellular processes in living animals.Methods. Following Our recent development of inbred green fluorescent protein (GFP)-transgenic (Tg) rats, we created the establishment of inbred (Lewis) Tg rats with firefly luciferase. The immunogenicity against luciferase was evaluated by the skin grafting test, and the fate of grafts was monitored by in vivo luminescent technique.Results. The luciferase-Tg rats ubiquitously expressed the marker gene. Conventional skin grafting apparently showed the long-term acceptance of luciferase-Tg rat skin on wild-type rats (> 100 days), compared with grafting of GFP-Tg-derived skin (< 10 days). This Suggests less cellular immune responsiveness against the luciferase protein than GFP. Strikingly, organ transplants with heart and small bowel, and bone marrow cell transplantation showed viability and graft acceptance, demonstrating that cells and organs from luciferase-Tg rats are transplantable and their fate can be tracked for a sufficient time. Taking advantage of less immunogenic luciferase, cellular fate of transplanted mature hepatocytes was also examined. Transplanted hepatocytes proliferated and were monitored selectively in damaged liver, but not in healthy liver, for over 60 days.Conclusions. We propose on the basis of these findings that the luciferase-Tg rat system with modern optical imaging offers a new platform for a better understanding of stem cell biology and transplantation.