Biologists at the University of Sheffield’s Center for Stem Cell Biology have completed a landmark study of human pluripotent stem cells. The findings were published this week in the journalStem Cell Reports. The researchers used time-lapse imaging of single human embryonic stem cells to observe their behavior and identify factors that restrict growth as well as factors that enable cells to grow more efficiently.
Currently cells tend to die extensively during culturing and they can mutate spontaneously. Some of these genetic mutations are known to provide stem cells with superior growth capabilities, enabling them to overtake the culture – a phenomenon termed culture adaptation, which mimics the behavior of cancer cells.
The team’s research combined the use of time-lapse microscopy, single-cell tracking and mathematical modelling to identify development stages that affect the survival of normal human embryonic stem cells and compared them with adapted cells. They identified three major bottlenecks affecting colony formation: survival after plating, failure to re-enter into cell cycle and continued cell death after division. In the same culture condition, they found adapted cells performed better in all of these points leading to more colonies. Bottlenecks were also alleviated through cell to cell contact and pro-survival compounds.
Veronica Biga, PhD, from the University’s Automatic Control and Systems Engineering Department, says: “To extract information about cell death, mitosis and movement, we developed new methods for analyzing images and measuring numerous parameters from time-lapse videos.”
She adds: “We plan to further develop the methods from this study into an image processing and analysis software solution to be used for monitoring cell behavior in applications such as screening culture conditions, drug discovery, monitoring and minimizing the occurrence of genetic abnormalities directly through time-lapse.” Read the study abstract.
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