Enzyme-free release of adhered cells from standard culture dishes using intermittent ultrasonic traveling waves
Enzyme-free release of adhered cells from standard culture dishes using intermittent ultrasonic traveling waves
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DOI：
10.1038/s42003-019-0638-5
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发表时间：
2019-10-29
影响因子：
5.9
通讯作者：
Takemura, Kenjiro
中科院分区：
文献类型：
--
作者：
Kurashina, Yuta;Imashiro, Chikahiro;Takemura, Kenjiro
在培养贴壁细胞时，细胞分离是必不可少的。胰酶消化是最常用的脱离技术，尽管它会由于膜和细胞外基质的破坏而降低存活率。避免这种损害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，在无血清介质中从间歇性行波中晃动。该方法的细胞分离率为96.2%，48h的转移率为常规方法的130%，与胰酶消化法分离的细胞数相比，其转移率提高了130%。我们发现，消除胰酶可以减少细胞损伤，提高分离细胞的存活率。施加在细胞上的声压和间歇性行波引起的介质晃动是导致细胞脱落的最重要因素。这种建议的方法将通过更有效的转移过程加快组织培养细胞的扩增，从而提高生物制药的生产。
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.