Escriou V, Ciolina C, Lacroix F, Byk G, Scherman D, Wils P
UMR 133 CNRS/Rhône-Poulenc Rorer, Centre de Recherche de Vitry-Alfortville, Vitry sur Seine, France.
Biochim Biophys Acta. 1998 Jan 19;1368(2):276-88. doi: 10.1016/s0005-2736(97)00194-6.
Most of the cationic lipids used for gene transfer experiments drastically lose their efficiency in the presence of serum. We used a cationic lipid with a spermine head group and its fluorescent analog to study the cellular uptake and the intracellular fate of lipoplexes in the presence and absence of serum. We found that the amount of DNA and lipid taken up by the cells was not related to the efficacy of the gene transfer. When the lipofection was performed in the presence of serum, lipoplexes were contained within small intracellular vesicles. In the absence of serum, the vesicles were larger and heterogeneous in size and shape. By analysis of their size distribution, we showed that lipoplexes preformed in the absence of serum tended to aggregate. This aggregation was inhibited in the presence of serum. We used a carbonate formulation that led to the preformation of large particles: those large particles gave a high lipofection efficiency in the presence of serum and their intracellular distribution was identical to that observed in the absence of serum.
大多数用于基因转移实验的阳离子脂质在血清存在的情况下会大幅降低其效率。我们使用了一种带有精胺头部基团的阳离子脂质及其荧光类似物，来研究在有血清和无血清情况下脂质体的细胞摄取及细胞内命运。我们发现细胞摄取的DNA和脂质的量与基因转移的效率无关。当在血清存在的情况下进行脂质转染时，脂质体包含在小的细胞内囊泡中。在无血清的情况下，囊泡更大且大小和形状各异。通过对其大小分布的分析，我们表明在无血清条件下预先形成的脂质体倾向于聚集。这种聚集在血清存在时受到抑制。我们使用了一种能导致大颗粒预先形成的碳酸盐配方：那些大颗粒在血清存在时具有高脂质转染效率，并且它们在细胞内的分布与在无血清情况下观察到的相同。
