Bauer Clayton T, Shtridelman Yuri, Lema Tomé Carla M, Grim Joel Q, Turner Christopher P, Tytell Michael, Macosko Jed C
Physics Department, Wake Forest University, Winston-Salem, NC 27109-7507, USA.
Neurosci Lett. 2008 Aug 22;441(2):173-7. doi: 10.1016/j.neulet.2008.06.026. Epub 2008 Jun 13.
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.
由于观察单个细胞的便捷性和实用性，原代神经元培养物在研究中被广泛使用。因此，了解培养条件的变化如何影响神经元生理学至关重要。培养神经元的一个潜在变化是细胞内运输的改变。由于运输对于细胞器、蛋白质、核酸和脂质的正常递送是必需的，它是细胞生理学的一个合理指标。我们测试了这样一个假设，即细胞器运输可能会随着体外群体密度的变化而改变，从而表明细胞生理学的变化。使用一种新颖的背景减除成像方法，我们发现，在体外培养5天（DIV）时，胚胎大鼠脊髓神经元中囊泡细胞器的运输与细胞密度呈正相关。当密度增加6.5倍时，运输的细胞器数量增加了2.2±0.3倍。有趣的是，在3 - 4 DIV时未观察到这种效应。这些结果表明，在接种程序相对较小的变化下，细胞生理学发生了显著变化；这表明看似形态相似且处于相同DIV的细胞，可能仍然存在很大程度的变异性。
