Achieving Greater Efficiency and Higher Confidence in Single-cell Cloning by Combining Cell Printing and Plate Imaging Technologies

In recent years, health authorities have increased emphasis on demonstrating that a cell line, which is used for the generation of biologics, is clonally derived. Within the past few years, single-cell manipulation technologies, especially microfluidic drop-on-demand dispensing, have gained increased attention in the biopharmaceutical industry. This work discusses the development and characterization of a single-cell printing workflow followed by plate imaging. By combining single-cell printing and plate imaging with manual image verification it is possible to, (1) dramatically reduce the number of microtiter plates needed during the single-cell cloning of clinical cell lines, as compared with a limiting-dilution single-cell cloning workflow, and therefore reduce the number of high-resolution images acquired and stored and (2) achieve >99.99% assurance that the cell lines derived from this workflow are clonally derived. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 34:1454-1459, 2018.