Microcarrier based production of cryopreserved cells
Poster Nov 13, 2009
Sharon Davies, Lynne Smith, Sian Kalinka, Philip Meyler, Chris Jones, Rahman Ismail, Elizabeth Price, Stephen Game, J M Kendall., GE Healthcare
The use of cellular assays in drug screening continues to grow with over 50% of primary screens using cell based formats in 2006. The majority of cells used in the drug discovery industry are fresh, using ‘just in time’ batch processing from in house facilities. This process could give rise to a number of issues, namely batch variation, scheduling of cell production and capacity management. We describe the use of microcarrier technology in combination with a suitably configured bioreactor to provide a rapid, robust and reproducible approach to cell production. Cells cultured in this way can be removed from beads, cryopreserved in a single HTS batch size format and maintain assay capacity.
We utilized paired synthetic crRNAs coupled with our synthetic tracrRNA in cells transduced with lentiviral Cas9 to perform a functional knockout on hsa-miR-221. This three-part system (crRNA, tracrRNA and Cas9) has demonstrated efficient gene editing when used with only one guide RNA, but the goal was to use two crRNAs to remove the entire stem-loop.READ MORE