Monoclonal cell line generation from small-scale to high-throughput labs

Generating monoclonal cell lines is essential for a variety of scientific applications. From ensuring uniform gene expression in genetically engineered disease research lines to stabilizing production of antibodies for industrial manufacturers, there is a ubiquitous need for labs to isolate single cells for monoclonal cell line expansion. However, despite the demand for quick and cost-effective options, most labs still utilize the manual and inefficient cell cloning method known as limiting dilution (LD).

Manual limiting dilution methods limit productivity

According to Poisson distribution statistics,1 when cells are diluted and dispensed at 1 cell/well, 37% of wells should contain a single cell. More dilute concentrations can further eliminate doublets, with expected frequencies of 19% to 30% single-cell wells for 0.25 to 0.50 cells/well concentrations, respectively. These statistics are often inflated by the assumption that all viable clones have the same random chance of monoclonality,2 however, even under perfect conditions, 2/3 of each plate is still rendered useless. This wastes a large amount of supplies and time to produce sufficient monoclonal colony numbers.

Improve time, cost, and expression efficiency with the WOLF

The NanoCellect WOLF Cell Sorter significantly reduces all these resources wasted by the limiting dilution method. The WOLF and N1 single cell dispenser use sterile disposable microfluidic cartridges to analyze, sort, and dispense single cells into 96- or 384-well plates in as little as 8 minutes with over 90% efficiency.3 The WOLF is small enough to fit within a biosafety cabinet to maintain sterility; it provides 2 scatter and 3 fluorescent channels for selection of your cells of interest. Below, Chinese Hamster Ovarian (CHO) cells, the leading cell line used by industrial manufacturers for protein production, are used to demonstrate the substantially higher efficiency of the WOLF when compared with a range of limiting dilution concentrations.

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