A Simple Method for in situ Quantification of Cells on Carriers

Bio Protoc. 2021 Dec 5;11(23):e4254. doi: 10.21769/BioProtoc.4254.

Abstract

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 in situ quantification method.

Keywords: Alamar blue; Carriers; Cells; Fluorescence; High-throughput quantification; Plate reader; Quantification.