Alvarez F J, Herráez A, Tejedor M C, Díez J C
Departamento de Bioquímica y Biología Molecular, Universidad de Alcalá, Madrid, Spain.
Biotechnol Appl Biochem. 1996 Apr;23(2):173-9.
Rat and human carrier red blood cells (RBCs) were prepared by hypotonic-dialysis encapsulation. The standard conditions used for encapsulation were 80 mOsm/kg for 60 min. The encapsulation behaviour of rat and human RBCs was studied using radiolabelled carbonic anhydrase and fluorescently labelled dextran. Both markers are incorporated to slightly greater extents by human than by rat RBCs by hypotonic treatment. Cell recovery of rat and human RBCs loaded with either carbonic anhydrase or fluorescent dextran accounted for 49% and 80% respectively. The cellular integrity of the loaded cells was revealed by the presence of fluorescence labelling in rat and human RBCs. Fluorescence studies showed an increase of size dispersion in loaded rat and human RBCs, giving cellular volume variations in both types of cells resulting from the encapsulation procedure. Two loaded cell populations were evident in both species, one with high fluorescence content and another with background staining. Apparently the proportion of high fluorescently labelled loaded cells was higher in the case of the human RBCs. A reduced level of fluorescence labelling was observed in rat and human RBC membranes, which indicates a process of adsorption of dextran to the membranes.
通过低渗透析包封法制备大鼠和人载体红细胞（RBCs）。用于包封的标准条件为80 mOsm/kg，持续60分钟。使用放射性标记的碳酸酐酶和荧光标记的葡聚糖研究大鼠和人红细胞的包封行为。通过低渗处理，两种标记物在人红细胞中的掺入程度均略高于大鼠红细胞。负载碳酸酐酶或荧光葡聚糖的大鼠和人红细胞的细胞回收率分别为49%和80%。通过大鼠和人红细胞中荧光标记的存在揭示了负载细胞的细胞完整性。荧光研究表明，负载的大鼠和人红细胞的大小分散性增加，包封过程导致两种类型的细胞出现细胞体积变化。在两个物种中都明显存在两个负载细胞群体，一个具有高荧光含量，另一个具有背景染色。显然，人红细胞中高荧光标记的负载细胞比例更高。在大鼠和人红细胞膜中观察到荧光标记水平降低，这表明葡聚糖吸附到膜上的过程。
