Application Note
Automation improves development of stem cell cultures
Advantages of automation in stem cell culturing
Introduction
Material and methods
Stem cells are found in all multicellular organisms, and are
characterized by their ability to differentiate into a diverse
range of specialized cell types. One of the most important
potential applications of human stem cells is cell-based
therapy, where cells and tissues could be generated and used
to treat many diseases such as Alzheimer’s disease, spinal
cord injury and stroke. Another application for human stem
cells is for the testing of new drugs in biopharmaceutical
development. To screen and compare different drugs
effectively, the testing conditions, including the quality of the
cells, must be identical. It is therefore essential to precisely
control the differentiation of stem cells into the specific cell
type, on which the drugs are intended to be tested. However,
stem cells are difficult to grow with good reproducibility; high
variability and poor consistency of cultures present a
challenge for repeatable experimental protocols in stem cell
research. Automation is an obvious means for solving these
problems, improving the consistency of stem cell culturing by
performing every step identically and under controlled
conditions, maintaining cultures for precise, predetermined
durations.
Prof Gary Lye and Dr Farlan Veraitch at the Department of
Biochemical Engineering at University College London (UCL),
UK, have set up a system for the culture of stem cells in 24well plates. The focus of the research was on the reliable
production of pluripotent stem cell populations as well as the
differentiation of the stem cells into specific cell types, such as
neuronal cells. Manual and automated methods of culturing
pluripotent stem cell populations were compared through
several passages, involving multiple operations including
media exchanges and re-plating of the cells. The automated
®
system consisted of a Tecan Freedom EVO liquid handling
platform, which was installed in a biological safety cabinet to
guarantee both operator safety and protection of the stem cell
cultures from contamination. The cabinet was temperaturecontrolled, and levels of CO2 and O2 were also automatically
regulated. Plates were stored in an automated CO2 incubator
placed at the right side of the workstation to ensure that the
cells were not subjected to variation in CO2 levels, and
consequently pH, during transfer between the CO2 incubator
and the Freedom EVO deck.
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Application Note
Results
Fig. 1 Automated stem cell growth workstation at UCL.
The system comprises of the following components:

Freedom EVO 100 (Tecan)

4-channel liquid handling arm (LiHa) with low volume
option, using disposable tips (DiTi)

robotic manipulator arm with extended reach (RoMa long)
and centric gripper to load plates into the centrifuge

Centrifuge (Rotanta 46RSC, Hettich® GmbH) for
centrifugation of cells in microplates

Automated CO2 incubator (Cytomat, Thermo Fisher
Scientific) for incubation of stem cells in microplates

Safety cabinet (Walker Safety Cabinets) with controlled
environment for O2, CO2 and temperature
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Stem cells from a variety of sources could be grown in 24-well
plates as pluripotent stem cells, or differentiated into specific
cell types, such as neuronal cells. Following expansion, stem
cells retained the ability to differentiate into all three germ cell
types – neuroectoderm, mesoderm and endoderm – so, in
principle, any cell type could be accessed. Stem cell culture
and differentiation of neuronal cells was successfully
automated, and would potentially apply to any screening
process requiring consistent cell numbers and quality.
Fig. 3 Reproducible automated passaging of embryonic stem cells
Automation significantly reduced variability in pluripotencyspecific gene expression profiles, maintaining a more uniform
stem cell population. In addition, after differentiation into
neuronal cells, the cells cultured using the automated system
gave higher and more consistent levels of neuronal surface
markers and neuron-specific gene expression.
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Fig. 2 Detailed view on the automated stem cell growth workstation
Set up of the worktable (from left to right)
1. DiTi carrier
2. wash station with DiTi waste chute
3. heated carrier for 2× 250 ml and 5× 100 ml troughs
4. tilting rack with heated microplate holders
®
5. VARIOMAG Teleshake (Thermo Fisher Scientific)
Fig. 4 Immunocytochemistry of embryonic stem cells after 192 hours of
neural differentiation. Single staining for bIII tubulin-FITC (green).
Neuronal cells were identified in clusters showing typical neural rosette
morphology. DAPI (blue) shows cell nuclei
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Application Note
Conclusions
Acknowledgements
Automation has been able to provide stem cell cultures of
higher consistency and reproducibility compared to manual
methods. The risk of contamination during automated
handling has been minimized by transferring culture plates
directly between the incubator and worktable, without
exposure to airborne contaminants in the laboratory.
Maintaining the cultures within a closed system also improves
operator safety.
Professor Lye and Dr Veraitch would like to acknowledge Dr
Waqar Hussain and Mr Paul Mondragon-Teran who
performed the work described here, and also the contributions
of UCL colleagues Professor Peter Dunnill, Professor Chris
Mason and Dr Ivan Wall.
The results have demonstrated the potential advantages of
automated stem cell culture in biopharmaceutical and cellbased therapy applications, where a consistent quality of
cultured cells is essential.
For further information on UCL, visit
www.ucl.ac.uk/biochemeng/industry/regenmed
For further information on Tecan’s stem cell
solutions, visit www.tecan.com/stemcell
This scientific instrumentation is not for use in human clinical
or diagnostic procedures.
Tecan Group Ltd. makes every effort to include accurate and up-to-date information within this publication; however, it is possible that omissions or errors might
have occurred. Tecan Group Ltd. cannot, therefore, make any representations or warranties, expressed or implied, as to the accuracy or completeness of the
information provided in this publication. Changes in this publication can be made at any time without notice. All mentioned trademarks are protected by law.
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396293 V1.0 03-2010
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