Boehm Daniela, Bell Angus
Department of Microbiology, School of Genetics & Microbiology, Moyne Institute of Preventive Medicine, Trinity College Dublin, Dublin 2, Ireland.
Biotechnol Rep (Amst). 2014 Jul 24;4:34-41. doi: 10.1016/j.btre.2014.07.005. eCollection 2014 Dec.
Most mammalian cell proliferation assays rely on manual or automated cell counting or the assessment of metabolic activity in colorimetric assays, with the former being either labor and time intensive or expensive and the latter being multistep procedures requiring the addition of several reagents. The proliferation of erythroid cells from hematopoietic stem cells and their differentiation into mature red blood cells is characterized by the accumulation of large amounts of hemoglobin. Hemoglobin concentrations are easily quantifiable using spectrophotometric methods due to the specific absorbance peak of the molecule's heme moiety between 400 and 420 nm. Erythroid proliferation can therefore be readily assessed using spectrophotometric measurement in this range. We have used this feature of erythroid cells to develop a simple erythroid proliferation assay that is minimally labor/time- and reagent-intensive and could easily be automated for use in high-throughput screening. Such an assay can be a valuable tool for investigations into hematological disorders where erythropoiesis is dysregulated, i.e., either inhibited or enhanced, into the development of anemia as a side-effect of primary diseases such as parasitic infections and into cyto-(particularly erythro-) toxicity of chemical agents or drugs.
大多数哺乳动物细胞增殖试验依赖于手动或自动细胞计数，或比色法中代谢活性的评估，前者既耗费人力和时间，又成本高昂，而后者是需要添加多种试剂的多步骤程序。造血干细胞中红细胞的增殖及其向成熟红细胞的分化，其特征是大量血红蛋白的积累。由于该分子血红素部分在400至420纳米之间有特定的吸收峰，因此使用分光光度法很容易定量血红蛋白浓度。因此，在此范围内使用分光光度测量可以很容易地评估红细胞增殖。我们利用红细胞的这一特性开发了一种简单的红细胞增殖试验，该试验所需人力/时间和试剂最少，并且可以很容易地实现自动化以用于高通量筛选。这样的试验对于研究造血失调的血液疾病（即红细胞生成受到抑制或增强）、作为寄生虫感染等原发性疾病副作用的贫血症的发展以及化学试剂或药物的细胞毒性（特别是红细胞毒性）而言，可能是一种有价值的工具。
