Monitoring and analysis of dynamic growth of human embryonic stem cells: comparison of automated instrumentation and conventional culturing methods.
BACKGROUND
Human embryonic stem cells (hESCs) are a potential source of cells for use in regenerative medicine. Automation of culturing, monitoring and analysis is crucial for fast and reliable optimization of hESC culturing methods. Continuous monitoring of living cell cultures can reveal more information and is faster than using laborious traditional methods such as microscopic evaluation, immunohistochemistry and flow cytometry.


METHODS
We analyzed the growth dynamics of two hESC lines HS237 and HS293 in a conventional culture medium containing serum replacement and a xeno-free X-vivo 10 medium. We used a new automated culture platform utilizing machine vision technology, which enables automatic observation, recording and analysis of intact living cells. We validated the results using flow cytometry for cell counting and characterization.


RESULTS
In our analyses, hESC colony growth could be continuously monitored and the proportion of undifferentiated cells automatically analyzed. No labeling was needed and we could, for the first time, perform detailed follow up of live, undisturbed cell colonies, and record all the events in the culture. The growth rate of the hESCs cultured in X-vivo 10 medium was significantly lower and a larger proportion of the cells were differentiated.


CONCLUSIONS
The new automated system enables rapid and reliable analysis of undifferentiated growth dynamics of hESCs. We demonstrate the effectiveness of the system by comparing hESC growth in different culture conditions.
Narkilahti
Susanna
S
REGEA, Institute for Regenerative Medicine, University of Tampere and Tampere University Hospital, 33520 Tampere, Finland. Susanna.Narkilahti@regea.fi
Rajala
Kristiina
K
Pihlajamäki
Harri
H
Suuronen
Riitta
R
Hovatta
Outi
O
Skottman
Heli
H
D001185:Artificial Intelligence; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods; D049109:Cell Proliferation; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000502:physiology*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D006801:Humans; D007090:Image Interpretation, Computer-Assisted / Q000295:instrumentation / Q000379:methods; D007700:Kinetics; D018613:Microscopy, Confocal / Q000295:instrumentation* / Q000379:methods; D012371:Robotics / Q000295:instrumentation* / Q000379:methods