Higashi N, Sunamoto J
Supermolecules Project, Research Development Corporation of Japan, Kyoto.
Biochim Biophys Acta. 1995 Apr 13;1243(3):386-92. doi: 10.1016/0304-4165(94)00163-r.
Egg PC liposome as reconstituted with poly(ethylene oxide)-bearing lipid (coded as PEO-lipid (n = 15)) was remarkably endocytosed by Jurkat cell, which was a lymphoblastoma derived from human T cell. To confirm the endocytosis, two kinds of fluorescent probes (FITC-dextran and octadecyl rhodamine B) were employed. The former was loaded in the aqueous phase of the liposome, while the latter was embedded in the liposomal membrane. Both probes were found coincidentally at the same site in the cytosol, clearly suggesting that whole liposome entered the cell. The endocytosis was most obvious when PEO-lipid (n = 15) was employed above 50 mol%. FITC-Dextran entered the cell was found small dots in the cell, not dispersive. Even when octadecyl rhodamine B was used, no membrane fluorescence was observed at all. The uptake closely related to the cell metabolism as affected by the culture temperature and serum in the incubation medium. Furthermore, the addition of cytochalasin B completely prohibited the cell uptake of liposomes.
用含聚环氧乙烷脂质(编码为聚环氧乙烷脂质,n = 15)重构的鸡蛋磷脂酰胆碱脂质体被人T细胞来源的淋巴瘤细胞Jurkat细胞显著内吞。为了证实内吞作用,使用了两种荧光探针(异硫氰酸荧光素 - 葡聚糖和十八烷基罗丹明B)。前者加载到脂质体的水相中,而后者嵌入脂质体膜中。两种探针都在细胞质中的同一位置同时被发现,清楚地表明整个脂质体进入了细胞。当聚环氧乙烷脂质(n = 15)的使用比例高于50摩尔%时,内吞作用最为明显。进入细胞的异硫氰酸荧光素 - 葡聚糖在细胞中呈小点状,而非分散状。即使使用十八烷基罗丹明B,也完全没有观察到膜荧光。摄取与培养温度和孵育培养基中的血清影响的细胞代谢密切相关。此外,加入细胞松弛素B完全抑制了脂质体的细胞摄取。