Most cell biology, biomaterials and associated research is conducted on  cells attached to tissue culture plastic in multiwell plates - such as  high throughput drug discovery and toxicity, to viral plaque assays.  However, there is a disconnect that the cells are stored frozen in  suspension, not in the format ‘ready to use’. This is because  conventional cryoprotectants do not protect the cells when in monolayer format. 

The GibsonGroup, and UoW Spin-Out Cryologyx have worked together  to solve this problem using macromolecular cryoprotectants. In this  later paper, the team demonstrate reproducible and robust recovery of  cell monolayers out of the freezer. This is shown for common cell lines, including HepG2 and Caco-2, commonly used in drug screening. This is a  revolutionary technology as it shows researchers could stop wasting time culturing cells, and just order them, remove from freezer and within 24  hours begin data collection with no of the traditional culturing steps. Cryologyx are deploying these findings to commercialise assay ready  cells, and trial plates are available!

Read the abstract below, or the full paper here

Cell monolayers underpin the discovery and screening of new drugs and  allow for fundamental studies of cell biology and disease. However,  current cryopreservation technologies do not allow cells to be stored  frozen while attached to tissue culture plastic. Hence, cells must be  thawed from suspension, cultured for several days or weeks, and finally  transferred into multiwell plates for the desired application. This  inefficient process consumes significant time handling cells, rather  than conducting biomedical research or other value-adding activities.  Here, we demonstrate that a synthetic macromolecular cryoprotectant  enables the routine, reproducible, and robust cryopreservation of  biomedically important cell monolayers, within industry-standard tissue  culture multiwell plates. The cells are simply thawed with media and  placed in an incubator ready to use within 24 h. 

Post-thaw cell recovery  values were >80% across three cell lines with low well-to-well  variance. The cryopreserved cells retained healthy morphology, membrane  integrity, proliferative capacity, and metabolic activity; showed  marginal increases in apoptotic cells; and responded well to a  toxicological challenge using doxorubicin. These discoveries confirm  that the cells are “assay-ready” 24 h after thaw. Overall, we show that  macromolecular cryoprotectants can address a long-standing  cryobiological challenge and offers the potential to transform routine  cell culture for biomedical discovery.

© 2022 The Authors. Published by American Chemical Society