1Department of Mechanical Engineering, Faculty of Science and Technology, Keio University, 3-14-1 Hiyoshi, Kohoku-ku, Yokohama 223-8522 Japan.
2Department of Materials Science and Engineering, School of Materials and Chemical Technology, Tokyo Institute of Technology, 4259 Nagatsutacho, Midori-ku, Yokohama, 226-8503 Japan.
Commun Biol. 2019 Oct 29;2:393. doi: 10.1038/s42003-019-0638-5. eCollection 2019.
Cell detachment is essential in culturing adherent cells. Trypsinization is the most popular detachment technique, even though it reduces viability due to the damage to the membrane and extracellular matrix. Avoiding such damage would improve cell culture efficiency. Here we propose an enzyme-free cell detachment method that employs the acoustic pressure, sloshing in serum-free medium from intermittent traveling wave. This method detaches 96.2% of the cells, and increases its transfer yield to 130% of conventional methods for 48 h, compared to the number of cells detached by trypsinization. We show the elimination of trypsinization reduces cell damage, improving the survival of the detached cells. Acoustic pressure applied to the cells and media sloshing from the intermittent traveling wave were identified as the most important factors leading to cell detachment. This proposed method will improve biopharmaceutical production by expediting the amplification of tissue-cultured cells through a more efficient transfer process.
细胞分离对于培养贴壁细胞是必不可少的。胰蛋白酶消化法是最常用的分离技术，但由于细胞膜和细胞外基质的损伤，它会降低细胞活力。避免这种损伤将提高细胞培养效率。在这里，我们提出了一种无酶细胞分离方法，该方法利用声压，在无血清培养基中通过间歇行波晃动来实现。与胰蛋白酶消化法相比，该方法可分离 96.2%的细胞，并且在 48 小时内将细胞转移产量提高到传统方法的 130%。我们表明，消除胰蛋白酶消化可减少细胞损伤，提高分离细胞的存活率。施加到细胞和介质中的声压以及间歇行波引起的介质晃动被确定为导致细胞分离的最重要因素。该方法将通过更有效的转移过程来加速组织培养细胞的扩增，从而提高生物制药的产量。
