Fibach E
Department of Hematology, Hadassah University Hospital, Jerusalem, Israel.
Hemoglobin. 1993 Feb;17(1):41-53. doi: 10.3109/03630269308998884.
The present paper reports on the application of a simple micromethod for quantitation of total and fetal hemoglobin in small volumes of diluted erythroid cell hemolysates. A modified alkaline denaturation procedure was used to denature and precipitate non-fetal hemoglobins and the tetramethylbenzidine procedure was used for quantitation of the soluble hemoglobin. Using a combination of these procedures, total hemoglobin concentrations as low as 0.2 mg% (2 micrograms/ml) and a fetal hemoglobin proportion of 0.5% could be reliably determined. We have utilized this procedure to study two aspects of hemoglobin production in human erythroid cells grown in culture; the proportion of fetal hemoglobin of cells in culture as compared to that in the peripheral blood of the donors, and the kinetics of hemoglobin production during maturation of these cultured cells. The results show that the cultured erythroid cells have a hemoglobin phenotype similar to the individual from whom they are derived and that erythroid differentiation may be studied in vitro in these cultured cells by measurement of these and other parameters.
本文报道了一种简单的微量方法在定量稀释的红系细胞溶血产物小体积样本中总血红蛋白和胎儿血红蛋白方面的应用。采用改良的碱性变性程序使非胎儿血红蛋白变性并沉淀，使用四甲基联苯胺程序对可溶性血红蛋白进行定量。结合这些程序，可可靠地测定低至0.2mg%（2微克/毫升）的总血红蛋白浓度以及0.5%的胎儿血红蛋白比例。我们利用该程序研究了培养的人红系细胞中血红蛋白产生的两个方面；培养细胞中胎儿血红蛋白的比例与供体外周血中胎儿血红蛋白比例的比较，以及这些培养细胞成熟过程中血红蛋白产生的动力学。结果表明，培养的红系细胞具有与其来源个体相似的血红蛋白表型，并且通过测量这些及其他参数，可以在体外对这些培养细胞中的红系分化进行研究。
