eBook: Optimizing Cell Culture Techniques - 12

Figure 2. HepG2 seeding on a 96 well plate containing
BIOMIMESYS hepatocyte hydrogels using a 96 channel
head on a VIAFLO 96 or VIAFLO 384 pipette.
Figure 1. BIOMIMESYS synthesis and SEM observation of a
section of the final hydrogel product.
and 300 μl Sterile, sterile, filter GRIPTIPS are used.
The entire application is performed in a laminar
flow cabinet.
Step-by-step procedure
1. Cell seeding on BIOMIMESYS hydrogel
with the VIAFLO 96 and VIAFLO 384
Homogenize HepG2 cells and seed on the readyto-use
BIOMIMESYS Hepatocyte hydrogel. This
step takes less than two minutes
HepG2 cells are homogenized in an automation
friendly reagent reservoir (150 ml or 300 ml) by
aspiration and dispensing. This is achieved using
the Pipet/mix mode of the VIAFLO 96 and
VIAFLO 384. Then, cells are transferred into a 96
well plate containing the BIOMIMESYS Hepatocyte
hydrogel. Selecting the right height (Z-height
limit: 2.9 mm) and pipetting speed (set on 2) settings
on the VIAFLO 96 and VIAFLO 384 is essential
to guarantee homogenous seeding of the cells on
the hydrogel (see Figure 2). Cell medium (150 μl)
is added by setting the Z-height limit to 3.2 mm
and the pipetting speed to 2.
2. Chlorpromazine treatment of HepG2
cell line
Remove culture medium and add the hepatotoxicity-inducing
drug chlorpromazine
Carefully remove 100 μl of medium and add 100 μl
of the drug, using a pipetting speed of 1 to prevent
harming the cells. A Z-height limit of 3 mm is set
to ensure spheroids are not accidently aspirated or
destroyed by the pipette tip.
3. HepG2 viability assay
Determine the metabolic activity of HepG2 cells
using a WST-1 assay
Remove 100 μl of culture medium using the normal
Pipet mode at speed 2 and a Z-height limit of
3 mm. Add 10 μl of the WST-1 reagent using the
same pipette settings described in step 2.
12
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