High Efficiency Gentle Sorting Improves Outgrowth for Single-Cell Cloning of Mammalian Cells
Single-cell cloning is required for biological workflows such as production of antibodies, stem cell therapy, and gene editing. Colonies that do not arise from a single cell can
result in discrepancies in product quality, instability of recombinant proteins, and issues with cellular growth rates. Efficient cloning methods should be able to discriminate cells
of interest from a heterogenous population and ensure that it can deposit one cell per well to establish cell lines with single cell origin. In addition to selecting a cloning method,
cells require optimized cloning medium to ensure survival at such low densities by using formulations that mimic a greater number of cells to support cell outgrowth. The WOLF
and WOLF G2 Cell Sorters alongside the N1 Single Cell Dispenser rely on microfluidics technology to gently sort at a pressure of <2 psi, reduce shear stress, and maintain a
close system for favorable biosafety. Furthermore, 50 mL of customized sheath buffer can be used to prevent increase cell death during sorting.
The WOLF Cell Sorter with the N1 Single Cell Dispenser was used to sort suspension (CHO ES, Expi293F, and Jurkat) and adherent (HEK293, MCF-7, A549, CHO-K1) cell
lines at 1 cell/well and were grown up to day 14 in suggested optimal cloning medium. Dispensing with the WOLF was compared with limiting dilution to demonstrate increased
efficiency when using this platform for cell line development.