An injectable spheroid system with genetic modification for cell transplantation therapy
An injectable spheroid system with genetic modification for cell transplantation therapy
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DOI：
10.1016/j.biomaterials.2013.12.012
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发表时间：
2014-03-01
期刊：
影响因子：
14
通讯作者：
Kataoka, Kazunori
中科院分区：
文献类型：
--
作者：
Uchida, Satoshi;Itaka, Keiji;Kataoka, Kazunori
通过使用移植细胞的三维球状体，随后用非病毒DNA载体，复合物纳米胶束进行遗传修饰，开发了增加细胞移植治疗潜力的新方法。具体地，在用热敏聚合物包被的微图案化培养板上形成用荧光素酶基因转染的原代肝细胞的调节大小为100 μ m的球状体，并简单地通过冷却板以可注射液体悬浮液的形式回收。皮下移植这些肝细胞球状体后，在宿主组织中观察到有效的转基因表达一个多月，而移植单层培养物的单细胞悬浮液仅导致短暂表达。球体系统有助于保存宿主组织中移植肝细胞的固有功能，如白蛋白表达，从而具有表达转基因的高潜力。移植细胞的活体观察表明，来自球状体培养物的细胞倾向于定位在血管附近，为保存细胞功能提供了有利的微环境。此外，用表达红细胞生成素的DNA转染的球状体显示出比单层培养的细胞悬浮液显着更高的造血作用，这表明这种基因修饰的球状体移植系统在治疗应用中具有很高的潜力。(C)2013爱思唯尔有限公司保留所有权利。
The new methodology to increase a therapeutic potential of cell transplantation was developed here by the use of three-dimensional spheroids of transplanting cells subsequent to the genetic modification with non-viral DNA vectors, polyplex nanomicelles. Particularly, spheroids in regulated size of 100-mu m of primary hepatocytes transfected with luciferase gene were formed on the micropatterned culture plates coated with thermosensitive polymer, and were recovered in the form of injectable liquid suspension simply by cooling the plates. After subcutaneously transplanting these hepatocyte spheroids, efficient transgene expression was observed in host tissue for more than a month, whereas transplantation of a single-cell suspension from a monolayer culture resulted in an only transient expression. The spheroid system contributed to the preservation of innate functions of transplanted hepatocytes in the host tissue, such as albumin expression, thereby possessing high potential for expressing transgene. Intravital observation of transplanted cells showed that those from spheroid cultures had a tendency to localize in the vicinity of blood vessels, making a favorable microenvironment for preserving cell functionality. Furthermore, spheroids transfected with erythropoietin-expressing DNA showed a significantly higher hematopoietic effect than that of cell suspensions from monolayer cultures, demonstrating high potential of this genetically-modified spheroid transplantation system for therapeutic applications. (C) 2013 Elsevier Ltd. All rights reserved.