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An injectable spheroid system with genetic modification for cell transplantation therapy

机译:具有遗传修饰的可注射球体系统,用于细胞移植治疗

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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-μm 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.
机译:通过在非病毒DNA载体多聚体纳米胶束进行基因修饰后,通过使用移植细胞的三维球体,开发了增加细胞移植治疗潜力的新方法。特别地,在用热敏性聚合物包被的微图案培养板上形成以大小为100μm的荧光素酶基因转染的原代肝细胞大小调节的球体,并通过冷却将其以注射液悬浮液的形式回收。皮下移植这些肝细胞球体后,在宿主组织中观察到有效的转基因表达已超过一个月,而从单层培养物中移植单细胞悬液仅导致瞬时表达。球体系统有助于保留宿主组织中移植的肝细胞的固有功能,例如白蛋白表达,从而具有表达转基因的高潜力。对移植细胞的活体内观察表明,来自球状培养物的细胞倾向于在血管附近定位,为保持细胞功能性提供了有利的微环境。此外,用表达促红细胞生成素的DNA转染的球体显示的造血作用要比单层培养物中的细胞悬浮液显着更高,这表明该基因修饰的球体移植系统具有很高的治疗潜力。

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