Pérez M T, Alvarez F J, García-Pérez A I, Lucas L, Tejedor M C, Sancho P
Departmento de Bioquímica y Biología Molecular, Universidad de Alcalá, Madrid, Spain.
J Chromatogr B Biomed Appl. 1996 Feb 23;677(1):45-51. doi: 10.1016/0378-4347(95)00433-5.
Carrier rat erythrocytes loaded with exogenous substances ([125I] carbonic anhydrase) by hypotonic-isotonic dialysis become heterogeneous cell populations that can be fractionated using the counter-current distribution (CCD) technique. Two well-defined low- and high-partition ratio, G, subpopulations are obtained in charge-sensitive dextran-polyethylene glycol two-phase systems. The low-G subpopulation, which contains the most fragile and surface-altered cells, as deduced from their osmotic fragility curves and partition behaviour, respectively, presents a high amount of exogenous substance incorporated (134.6 cpm/10(6) cells). The high-G subpopulation, that contains cells similar to the control or isotonically dialyzed cells presents a lower amount of exogenous substance incorporated (69.8 cpm/10(6) cells). Cells in this high-G subpopulation seem to be fractionated, like the controls, according to ageing as suggested by the decline of the pyruvate kinase specific activity from the left- to the right-hand side of the CCD profile.
通过低渗-等渗透析加载外源物质（[125I]碳酸酐酶）的携带大鼠红细胞成为异质细胞群体，可使用逆流分配（CCD）技术进行分级分离。在电荷敏感的葡聚糖-聚乙二醇双相系统中可获得两个明确的低分配比和高分配比G亚群。从其渗透脆性曲线和分配行为分别推断，低G亚群包含最脆弱和表面改变的细胞，其掺入的外源物质含量较高（134.6 cpm/10(6)个细胞）。高G亚群包含与对照或等渗透析细胞相似的细胞，其掺入的外源物质含量较低（69.8 cpm/10(6)个细胞）。如CCD图谱从左侧到右侧丙酮酸激酶比活性的下降所示，该高G亚群中的细胞似乎像对照一样根据老化进行分级分离。
