eBook: Optimizing Cell Culture Techniques - 7

Precise and Reproducible
Automated Cell Passaging Protocol
Mouse embryonic stem cell passaging with VIAFLO 96 and VIAFLO
384 handheld electronic pipettes
Cell culture is a technique in which cells are grown
outside a living organism under well-defined, controlled
conditions. It is important for basic biological
research and a wide range of clinical studies.
Mass culture of cells is indispensable for producing
viral vaccines, enzymes, synthetic hormones,
anticancer agents, recombinant proteins and antibodies,
but also for studying the physiology and
biochemistry of cells. Nowadays, cell culture is also
used in agriculture for producing alternative foods.
Preserving cell viability, avoidance of contamination,
high experimental reproducibility, and accuracy
are vital for manipulations carried out on cells
during media changes, transfers, seeding and passaging.
All these can be mastered with VIAFLO 96
and VIAFLO 384 handheld electronic pipettes.
Key benefits
* This protocol is automated for maximum
reproducibility. Reducing the hands-on time
for laboratory personnel can help to diminish
the risk of contamination.
* The optimized Z-height and mixing step setup
eliminates user variability. This is crucial
to maximize viability during cell culture.
* Pipetting is carried out at an accurate and
consistent speed to minimize the shearing
forces that cells are subjected to.
* The VIAFLO 96 and VIAFLO 384 combine
all the benefits of VIAFLO electronic
multichannel pipettes with the ability to
pipette 24, 96 or 384 handheld electronic
pipettes channels simultaneously, for
unmatched throughput and precision.
Overview: mouse embryonic stem
cell passaging protocol
Experimental set-up
In this protocol, ES-E14TG2 mouse embryonic stem
cells (Sigma) are washed, harvested and transferred
to a new plate with fresh media, using a VIAFLO 384
handheld electronic pipette with a three position
stage (with positions A, A/B and B, Figure 1). For the
aspiration steps, a VACUBOY vacuum hand operator
is used in combination with a VACUSAFE aspiration
system.
A 96 channel pipetting head (10-300 μl) is used together
with 300 μl sterile, filter GRIPTIPS® pipette
7
eBook: Optimizing Cell Culture Techniques

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