Stem Cell Research – Consistent Cell Counting for …

Apart from culture media, surface coatings, and pluripotency level of the cells you work with, the cell density is the fourth major contributor to experimental success. Intercellular signaling is difficult to control, but seeding cells at the right concentration can significantly contribute towards getting the directed differentiation of interest. For instance, differentiation protocols designed for mesoderm and endoderm cell types (e.g. pancreatic progenitors) have shown that low cell densities positively affect percentages of the cells of interest1,2. As an added benefit, low cell densities mean lower cytokine concentrations are required to obtain the same effect, and in turn saving considerable costs at the cell culture step.

Conversely, neural differentiation protocols often require high cell densities, as cell-to-cell signaling is indispensable and cell density at protocol onset determines the ratio of neuronal cells and cerebral organoids3,4.

Maintenance cultures of hPSCs are usually carried out by passaging dense cultures as medium-sized aggregates (clumps) of cells instead of dissociating them to single cells before re-plating. This ensures that the hPSC niche is not disrupted too much during passage and over multiple passages, which could lead to genetic drift and a decrease of pluripotency.

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Stem Cell Research - Consistent Cell Counting for ...