Cell type Advantages Disadvantages
Erythrocytes
Small volume of blood (3-10\(\mu\)l) required so suitable for finger prick test and multiple sampling. Doesn’t require any blood pre-processing so less hands on time. Reticulocyte percentage (%RET) in whole blood can be analysed simultaneously as a measure of haematopoietic function. Can detect an accumulation of DNA damage from repeat dosing over a prolonged period; long term effect. Cannot confirm mutant phenotype by DNA sequencing. Unsuitable for cryopreservation and batching samples from remote locations. PIG-A mutant erythrocytes may be subject to complement mediated lysis.
Reticulocytes
May be able to observe mutant induction sooner post exposure than erythrocytes. Reticulocyte percentage (%RET) in whole blood can be analysed simultaneously as a measure of haematopoietic function. PIG-A mutant reticulocytes would not be sensitive to complement mediated lysis.
Proportion of reticulocytes is low in whole blood compared to erythrocytes so would require larger blood volume.
Granulocytes
May be able to observe mutant induction sooner post exposure than erythrocytes. Allows for mutation confirmation using DNA sequencing. Requires blood pre-processing and a more complex flow cytometry gating strategy. Short life span of 1-2 days provides a small window of opportunity to capture mutant cells.
Peripheral blood mononuclear cells
Allows for mutation confirmation using DNA sequencing.
Published methodology includes subculture of PBMCs under Aerolysin selection. Requires larger volume of blood. Current studies show higher background level of mutant cells compared to erythrocytes.