2134/25121
Akinlolu O.O. Odeleye
Akinlolu O.O.
Odeleye
Sara Castillo-Avila
Sara
Castillo-Avila
Mathew Boon
Mathew
Boon
Haydn Martin
Haydn
Martin
Karen Coopman
Karen
Coopman
Development of an optical system for the non-invasive tracking of stem cell growth on microcarriers
Loughborough University
2017
untagged
Chemical Engineering not elsewhere classified
2017-05-25 09:24:41
Journal contribution
https://repository.lboro.ac.uk/articles/journal_contribution/Development_of_an_optical_system_for_the_non-invasive_tracking_of_stem_cell_growth_on_microcarriers/9242606
The emergence of medicinal indications for stem cell
therapies has seen a need to develop the manufacturing capacity for
adherent cells such as mesenchymal stem cells (MSCs). One such
development is in the use of microcarriers, which facilitate enhanced
cell densities for adherent stem cell cultures when compared with 2D
culture platforms. Given the variety of stem cell expansion systems
commercially available, novel methods of non-invasive and automated
monitoring of cell number, confluence, and aggregation, within
disparate environments, will become imperative to process control,
ensuring reliable and consistent performance. The in situ epiillumination
of mouse embryonic fibroblasts and human mesenchymal
stem cells attached to Cytodex 1 and 3 microcarriers was achieved using
a bespoke microscope. Robust image processing techniques were
developed to provide quantitative measurements of confluence,
aggregate recognition, and cell number, without the need for fluorescent
labeling or cell detachment. Large datasets of cells counted on individual
microcarriers were statistically analyzed and compared with NucleoCounter
measurements, with an average difference of less than 7%
observed from days 0 to 6 of a 12-day culture noted, prior to the onset of
aggregation. The developed image acquisition system and postprocessing
methodologies were successfully applied to dynamically
moving colonized microcarriers. The proposed system offers a novel
method of cell identification at the individual level, to consistently and
accurately assess viable cell number, confluence, and cell distribution,
while also minimizing the variability inherent in the current invasive
means by which cells adhered to microcarriers are analyzed.