Intraneuronal vesicular organelle transport changes with cell population density in vitro

Primary neuron cultures are widely used in research due to the ease and usefulness of observing individual cells. Therefore, it is vital to understand how variations in culture conditions may affect neuron physiology. One potential variation for cultured neurons is a change in intracellular transport. As transport is necessary for the normal delivery of organelles, proteins, nucleic acids, and lipids, it is a logical indicator of a cell's physiology. We test the hypothesis that organelle transport may change with varying in vitro population densities, thus indicating a change in cellular physiology. Using a novel background subtraction imaging method we show that, at 5 days in vitro (DIV), transport of vesicular organelles in embryonic rat spinal cord neurons is positively correlated with cell density. When density increased 6.5-fold, the number of transported organelles increased 2.2 ± 0.3-fold. Intriguingly, this effect was not observable at 3–4 DIV. These results show a significant change in cellular physiology with a relatively small change in plating procedure; this indicates that cells appearing to be morphologically similar, and at the same DIV, may still suffer from a great degree of variability.