Belhaj Marwa, Annamalai Balasubramaniam, Parsons Nathaniel, Shuler Andrew, Potts Jay, Rohrer Bärbel
Department of Cell Biology, University of South Carolina.
Department of Ophthalmology, Division of Research, Medical University of South Carolina.
J Vis Exp. 2020 Mar 30(157). doi: 10.3791/60162.
Many current therapeutics under development for diseases of the posterior pole of the eye are biologics. These drugs need to be administered frequently, typically via intravitreal injections. Encapsulated cells expressing the biologic of choice are becoming a tool for local protein production and release (e.g., via long-term drug delivery). In addition, encapsulation systems utilize permeable materials that allow diffusion of nutrients, waste, and therapeutic factors into and out of cells. This occurs while masking the cells from the host immune response, avoiding the need for suppression of the host immune system. This protocol describes the use of alginate as a polymer in microencapsulation coupled with the electrospray method as a microencapsulation technique. ARPE-19 cells, a spontaneously arising human RPE cell line, has been used in long-term cell therapy experiments due to its lifetime functionality, and it is used here for encapsulation and delivery of the capsules to mouse eyes. The manuscript summarizes the steps for cell microencapsulation, quality control, and ocular delivery.
目前许多正在研发用于治疗眼后极疾病的疗法都是生物制剂。这些药物通常需要通过玻璃体内注射频繁给药。表达所选生物制剂的封装细胞正成为局部蛋白质产生和释放的一种工具(例如,通过长期药物递送)。此外,封装系统利用可渗透材料,使营养物质、废物和治疗因子能够进出细胞。这一过程在使细胞免受宿主免疫反应影响的同时发生,避免了抑制宿主免疫系统的需求。本方案描述了使用藻酸盐作为聚合物进行微封装,并结合电喷雾法作为微封装技术。ARPE - 19细胞是一种自发产生的人视网膜色素上皮细胞系,因其具有终生功能而被用于长期细胞治疗实验,在此用于封装胶囊并将其递送至小鼠眼部。该手稿总结了细胞微封装、质量控制和眼部递送的步骤。