Red cell volume can be accurately determined in sheep using a nonradioactive biotin label.
Red cell volume can be accurately determined in sheep using a nonradioactive biotin label.
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DOI：
10.1203/pdr.0b013e318183f119
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发表时间：
2008-11
影响因子：
3.6
通讯作者：
Widness JA
中科院分区：
文献类型：
--
作者：
Mock DM;Mock NI;Lankford GL;Burmeister LF;Strauss RG;Widness JA
绵羊已成为研究人类胎儿和新生儿红细胞生成和红细胞(RBC)动力学的有用动物模型。我们之前验证了永久性标记(14C)氰酸盐用于测量绵羊红细胞体积(RCV)的有效性。在这里，我们验证了红细胞生物素标记是一种测量绵羊RCV的非放射性方法，并期望它可以应用于人类婴儿的研究。用两种生物素标记定量技术同时测定RCV。首次使用~(125)I链霉亲和素对生物素标记红细胞上生物素标记的血液总浓度进行了定量。第二组与荧光素标记的亲和素孵育后，用流式细胞仪计数生物素标记的红细胞。使用两种生物素定量技术进行的RCV测量以(14C)氰酸盐和51Cr作为参考方法进行了验证。这两种生物素技术产生的RCV值与参考方法以及彼此的RCV值非常一致，产生的相关系数平均≥为0.93.在同一动物中的连续重复测量也与(14C)氰酸盐方法和彼此一致(平均差值<；10%)。这些结果确立了生物素标记的红细胞作为一种准确的方法来测量绵羊的RCV。这种生物素方法可以应用于模拟新生儿红细胞生成的研究。
The sheep has served as an informative animal model for investigation of human fetal and newborn erythropoiesis and red blood cell (RBC) kinetics. We previously validated the permanent label (14C)cyanate for measuring red cell volume (RCV) in sheep. Here we validate biotin labeling of RBCs as a nonradioactive method for measuring RCV in sheep with the anticipation that it can be applied in studies of human infants. The RCV was determined simultaneously using two techniques for quantitation of the biotin label. The first quantified total blood concentration of biotin label on biotin-labeled RBCs using (125I)streptavidin. The second enumerated biotin-labeled RBCs by flow cytometry after incubation with fluorescein-conjugated avidin. RCV measurements made using the two-biotin quantitation techniques were validated against both (14C)cyanate and 51Cr as reference methods. Both biotin techniques produced RCV values that agreed well with the reference methods and with each other, producing correlation coefficients averaging ≥ 0.93. Sequential repetitive measurements in the same animal also agreed with the (14C)cyanate method and each other (average difference < 10%). These results establish biotin-labeled RBCs as an accurate method for performing RCV measurements in sheep. This biotin method can be applied in studies that model neonatal erythropoiesis.