Demediuk P, Anderson D K, Horrocks L A, Means E D
In Vitro Cell Dev Biol. 1985 Oct;21(10):569-74. doi: 10.1007/BF02620887.
The most commonly used procedure to harvest cultured cells from petri dishes is to scrape the cells off the plates with a rubber or Teflon policeman. However, the results reported herein demonstrate that this technique, with its associated mechanical trauma, significantly perturbed cell membranes in neuronal-enriched cultures derived from the ventral half of fetal murine spinal cords. This is evidenced by liberation of free fatty acids and diglycerides, partial inhibition of Na+,K+-ATPase activity, and increased malondialdehyde production. Harvesting the cells by freezing, either on liquid nitrogen or dry ice, significantly attenuated these effects. This important observation indicates that mechanical manipulation of cultured cells during harvesting significantly affects subsequent biochemical analyses, particularly those associated with the cell membrane (e.g., membrane lipid metabolism and assay of intrinsic membrane enzymes).
从培养皿中收获培养细胞最常用的方法是用橡胶或聚四氟乙烯刮棒将细胞从培养板上刮下。然而，本文报道的结果表明，这种技术及其相关的机械损伤，在源自胎鼠脊髓腹侧半部分的富含神经元的培养物中显著扰乱了细胞膜。游离脂肪酸和甘油二酯的释放、Na +，K + -ATP酶活性的部分抑制以及丙二醛产量的增加都证明了这一点。通过在液氮或干冰上冷冻来收获细胞，可显著减轻这些影响。这一重要观察结果表明，收获过程中对培养细胞的机械操作会显著影响后续的生化分析，特别是那些与细胞膜相关的分析（例如，膜脂代谢和内在膜酶的测定）。
