4
Notes
1. It is recommended to autoclave the bioreactor vessels as close
as possible to the time they will be used to ensure sterility is
maintained.
2. The arms of the Alvetex® insert line up with slots in the PTFE
holder to ensure a stable fit with no rotation. When the insert is
correctly positioned it should sit flat with minimal movement
possible.
3. Growth of HepG2 models for greater time points can be
carried out, with a further week of culture possible without
further media changes. For longer periods than this the quan-
tity of cells can lead to deleterious effects of the large tissue
mass such as the formation of a necrotic core.
4. If samples are not going to be imaged immediately, they can be
stored for several weeks in the dark at 4 C. For optimum
results, it is recommended to image the samples as soon as
possible.
References
1. Jensen C, Teng Y (2020) Is it time to start
transitioning from 2D to 3D cell culture? Front
Mol Biosci 7:1–15
2. Hansmann
J, Egger
D, Kasper C (2018)
Advanced dynamic cell and tissue culture. Bio-
engineering 5(3):5–7
3. Knight E, Przyborski S (2015) Advances in 3D
cell culture technologies enabling tissue-like
structures to be created in vitro. J Anat 227
(6):746–756
4. Sonbol HS (2018) Extracellular matrix remodel-
ing in human disease. J Microsc Ultrastruct 6
(3):123–128
5. Nicolas J, Magli S, Rabbachin L, Sampaolesi S,
Nicotra F, Russo L (2020) 3D Extracellular
matrix mimics: fundamental concepts and role
of materials chemistry to influence stem cell fate.
Biomacromolecules 21(6):1968–1994
6. Ballermann BJ, Dardik A, Eng E, Liu A (1998)
Shear stress and the endothelium. Kidney Int
Suppl 54(67):100–108
7. Verkman AS, Dix JA (1984) Effect of unstirred
layers on binding and reaction kinetics at a mem-
brane surface. Anal Biochem 142(1):109–116
8. Darling NJ, Mobbs CL, Gonza´lez-Hau AL,
Freer M, Przyborski S (2020) Bioengineering
novel in vitro co-culture models that represent
the human intestinal mucosa with improved
Caco-2 structure and barrier function. Front
Bioeng Biotechnol 8:1–15
256
Henry W. Hoyle et al.