Roberts T, De Boni U, Sefton M V
Department of Chemical Engineering and Applied Chemistry, University of Toronto, Ontario, Canada.
Biomaterials. 1996 Feb;17(3):267-75. doi: 10.1016/0142-9612(96)85564-5.
A rat pheochromocytoma cell line (PC12) was encapsulated in a water-insoluble hydroxyethyl methacrylate-methyl methacrylate copolymer by interfacial precipitation from a polyethylene glycol 200 solution into phosphate-buffered saline. The resulting capsules (660 +/- 44 microns in diameter; 84 +/- 27 microns wall thickness) contained viable PC12 cells in a spheroidal arrangement, much like tumour spheroids, the latter grown on surfaces unsuitable for cell attachment. In these spheroids, the viable cells formed a band approximately 100 microns thick, surrounding an inner core of necrotic cells. A similar arrangement was seen 14, 28 and 42 days after encapsulation, with capsules maintained in an in vitro tissue culture environment; the annular ring was roughly constant in size, although the packing density appeared to increase over the 6 week observation period. During the first 4 weeks, when measurements were made the encapsulated cells converted a tetrazolium dye (MTT) into an insoluble formazan product, in a time-after-encapsulation-dependent manner. This indicated that PC12 cells retained viability despite encapsulation and an ability to increase (at least in part) their metabolic capacity, presumably by a combination of proliferation and altered cellular activity. The encapsulated PC12 cells also secreted dopamine when incubated in a high potassium release medium but not in a low potassium, conventional tissue culture medium (RPMI 1640). Consistent with the MTT results, the amount of dopamine released was also dependent on the time after encapsulation, as well as the cell density at the time of encapsulation.
通过将聚乙二醇200溶液与磷酸盐缓冲盐水进行界面沉淀,将大鼠嗜铬细胞瘤细胞系(PC12)包封在水不溶性甲基丙烯酸羟乙酯-甲基丙烯酸甲酯共聚物中。所得胶囊(直径660±44微米;壁厚84±27微米)含有呈球状排列的活PC12细胞,很像肿瘤球体,后者生长在不适合细胞附着的表面上。在这些球体中,活细胞形成了一个约100微米厚的带,围绕着坏死细胞的内核。在包封后14、28和42天,当胶囊维持在体外组织培养环境中时,观察到类似的排列;尽管在6周的观察期内堆积密度似乎增加,但环的大小大致恒定。在最初的4周内,当进行测量时,包封的细胞以包封后时间依赖性的方式将四唑盐染料(MTT)转化为不溶性的甲臜产物。这表明PC12细胞尽管被包封仍保持活力,并且具有增加(至少部分)其代谢能力的能力,推测是通过增殖和细胞活性改变的组合。当在高钾释放培养基中孵育时,包封的PC12细胞也分泌多巴胺,但在低钾的常规组织培养基(RPMI 1640)中则不分泌。与MTT结果一致,释放的多巴胺量也取决于包封后的时间以及包封时的细胞密度。