Rosen Osnat, Jayson Avital, Natan Niva, Epstein Eyal
Department of Biotechnology, Israel Institute for Biological Research, Ness Ziona 7410001, Israel.
Bio Protoc. 2021 Dec 5;11(23):e4254. doi: 10.21769/BioProtoc.4254.
The technology of cell carriers was developed as a response to the need for high cell density to enable higher production levels in cell-based production processes. To follow the production process, quantifying the number of cells on these carriers is required, as well as tracking their viability and proliferation. However, owing to various carriers' unique structures, tracking the cells is challenging using current traditional assays that were originally developed for monolayers of adherent cells. The current "gold standard" method is counting cell nuclei, which is tedious and counts both live and dead cells. A few other techniques have been developed, but they are all specific to a carrier type and involve specialized equipment. Here, we describe a broad ranging method for counting cells on carriers. The method is based on the Alamar blue dye, a well-known, common marker for cell activity. No separation of the cells from the carriers is needed, nor is any specialized equipment. The method is simple and rapid, and provides comprehensive details necessary for control of production processes in cells. This method can be easily implemented in any cell-based process and other unique platforms for measuring growth of cells. Graphic abstract: Schematic of the quantification method.
细胞载体技术的发展是为了满足对高细胞密度的需求，以便在基于细胞的生产过程中实现更高的产量。为了跟踪生产过程，需要对这些载体上的细胞数量进行量化，同时还要跟踪它们的活力和增殖情况。然而，由于各种载体的独特结构，使用最初为贴壁细胞单层开发的当前传统检测方法来跟踪细胞具有挑战性。当前的“金标准”方法是对细胞核进行计数，这很繁琐，而且会同时计数活细胞和死细胞。已经开发了其他一些技术，但它们都特定于某种载体类型，并且需要专门的设备。在这里，我们描述了一种用于对载体上的细胞进行计数的广泛方法。该方法基于阿拉玛蓝染料，这是一种众所周知的、常用的细胞活性标记物。无需将细胞与载体分离，也不需要任何专门的设备。该方法简单快速，并提供了细胞生产过程控制所需的全面详细信息。这种方法可以很容易地应用于任何基于细胞的过程以及其他用于测量细胞生长的独特平台。图形摘要：量化方法示意图。
