微胶囊中细胞活性的比色测定法。

Colorimetric assay for cellular activity in microcapsules.

作者信息

Uludag H, Sefton M V

机构信息

Department of Chemical Engineering and Applied Chemistry, University of Toronto, Ontario, Canada.

出版信息

Biomaterials. 1990 Nov;11(9):708-12. doi: 10.1016/0142-9612(90)90032-l.

DOI:10.1016/0142-9612(90)90032-l

PMID:2090308

Abstract

Cellular activity in microcapsules was determined by a simple colorimetric assay, based on the cellular transformation of a tetrazolium salt, 3-(4,5-dimethyl-thiazol-2-yl)-2,5- diphenyl-tetrazolium bromide, into an insoluble formazan which was quantified in a spectrophotometer. The results showed that when encapsulated Chinese hamster ovary fibroblasts were exposed to the tetrazolium salt containing tissue culture medium, the formazan crystals were formed inside the poly(hydroxyethyl methacrylate-methyl methacrylate) microcapsules; capsules containing no cells or dead cells formed no formazan. A detectable amount of formazan was readily obtained even from single capsules. Formazan production was dependent on the incubation time, but not on the amount of added reagent. Capsules from a high cell-density encapsulation (4 X 10(6) cells/ml) formed more formazan than capsules from a low cell-density (4 X 10(5) cells/ml) encapsulation, suggesting a positive correlation between the cell density and tetrazolium transformation in microcapsules. The tetrazolium assay indicated the maintenance of cellular activity but slow, if any, proliferation in microcapsules over a 2 wk testing period.

摘要

通过一种简单的比色测定法来确定微胶囊中的细胞活性，该方法基于一种四唑盐（3-(4,5-二甲基噻唑-2-基)-2,5-二苯基溴化四唑）向不溶性甲臜的细胞转化，然后在分光光度计中对甲臜进行定量。结果表明，当将包封的中国仓鼠卵巢成纤维细胞暴露于含有四唑盐的组织培养基中时，在聚（甲基丙烯酸羟乙酯-甲基丙烯酸甲酯）微胶囊内部形成了甲臜晶体；不含细胞或死细胞的胶囊则不形成甲臜。即使从单个胶囊中也很容易获得可检测量的甲臜。甲臜的产生取决于孵育时间，但不取决于添加试剂的量。高细胞密度包封（4×10⁶个细胞/毫升）的胶囊比低细胞密度（4×10⁵个细胞/毫升）包封的胶囊形成更多的甲臜，这表明微胶囊中的细胞密度与四唑转化之间存在正相关。四唑测定表明在为期2周的测试期内微胶囊中的细胞活性得以维持，但增殖缓慢（如果有增殖的话）。