Semi-automated vs. manual: Comparative study of cell culture counting methods using validation parameters

Abstract

Determining cell density and cell viability is fundamental for any cell cultivation process. In addition to the manual counting method using hemocytometers, (semi-)automated methods offer advantages such as lower variability and shortened analysis times. However, these methods should provide at least comparable results to the manual method, which is why a comparison of methods is essential. We conducted a dilution series experimental design according to ISO 20391-2:2019 and compared two cell counting methods based on validation parameters aligned with the ICH Q2(R1) guideline. Regarding specificity and linearity, the manual (hemocytometer) and semi-automated (Countstar BioTech(R)) method exhibited similar results in the two evaluated characteristics total cell density and cell viability of CHO-K1 cells. Regarding repeatability of determining total cell density, the semi-automated method achieved significant ( = 0.05) better results with average relative standard deviations of 9 %. Concerning repeatability of the cell viability measurement, no significant difference between the two methods were shown. These results show the suitabililty of the dilution series experimental design. For the applied example, they indicate that the investigated semi-automated method is an appropriate alternative to the manual method.