Optimize, Adapt and Overcome - 8

Optimize, Adapt, and Overcome: Tips from the Cell Line Development Front * Single-Cell Printing for Establishing Monoclonality

Figure 2. Three graphs compare the efficiency and viability of clones between use of
a single-cell printer (SCP) and limiting dilution.
(A) The efficiency of single-cell deposit on the SCP is 88% compared with 32%
with limiting dilution.
(B) The percent outgrowth (or viability) was 84% for SCP and 81.7%
for limiting dilution.
(C) The SCP is compatible with many cell types as shown by the percent viability
of CHO-K1, HEK-293, and L-929. (Results may vary for your specified cell type.)

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(SCP). The likelihood of observing wells with multiple cells is also decreased compared with limiting
dilution, as the method is not statistically based.
With the SCP, only 6% of wells contained more than
one cell, whereas 10% of wells had multiple cells
using limiting dilution performed at a seeding
density of 0.5 cells/well (Figure 2A).
Beyond improving single-cell deposition efficiency,
use of the SCP may also improve viability. At the
2017 Cell Line Development conference, it was
reported in one study that the use of the SCP
delivered a cell viability >90%, whereas limiting
dilution yielded roughly 70% viable clones.1 This
study is preliminary, but it is supported by the
authors' findings (Figure 2B). The authors' current
hypothesis is that the gentle handling of cells within
the cartridge, in combination with cell screening
based on size and morphology, is a likely source
for the increase in viability as it offers the ability to
exclude unhealthy or otherwise asymmetrical cells.
On average, the authors have observed 75% viability across a variety of commonly used cell lines
(Figure 2C).

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