Quantification of the morphological characteristics of hESC colonies

The maintenance of the undifferentiated state in human embryonic stem cells (hESCs) is critical for further application in regenerative medicine, drug testing and studies of fundamental biology. Currently, the selection of the best quality cells and colonies for propagation is typically performed by...

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Bibliographic Details
Published in:Scientific reports 2019-11, Vol.9 (1), p.17569-11, Article 17569
Main Authors: Orozco-Fuentes, Sirio, Neganova, Irina, Wadkin, Laura E, Baggaley, Andrew W, Barrio, Rafael A, Lako, Majlinda, Shukurov, Anvar, Parker, Nicholas G
Format: Article
Language:English
Subjects:
Cell Culture Techniques
Cell Cycle
Cell interactions
Cell Proliferation
Colonies
Computer applications
Embryo cells
Human Embryonic Stem Cells - cytology
Human Embryonic Stem Cells - ultrastructure
Humans
Image processing
Microscopy, Phase-Contrast
Morphology
Nucleoli
Physical characteristics
Regenerative medicine
Stem cells
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Summary:The maintenance of the undifferentiated state in human embryonic stem cells (hESCs) is critical for further application in regenerative medicine, drug testing and studies of fundamental biology. Currently, the selection of the best quality cells and colonies for propagation is typically performed by eye, in terms of the displayed morphological features, such as prominent/abundant nucleoli and a colony with a tightly packed appearance and a well-defined edge. Using image analysis and computational tools, we precisely quantify these properties using phase-contrast images of hESC colonies of different sizes (0.1-1.1 [Formula: see text]) during days 2, 3 and 4 after plating. Our analyses reveal noticeable differences in their structure influenced directly by the colony area [Formula: see text]. Large colonies (A > 0.6 mm ) have cells with smaller nuclei and a short intercellular distance when compared with small colonies (A  0.6 mm ) due to the proliferation of the cells in the bulk. This increases the colony density and the number of nearest neighbours. We also detect the self-organisation of cells in the colonies where newly divided (smallest) cells cluster together in patches, separated from larger cells at the final stages of the cell cycle. This might influence directly cell-to-cell interactions and the community effects within the colonies since the segregation induced by size differences allows the interchange of neighbours as the cells proliferate and the colony grows. Our findings are relevant to efforts to determine the quality of hESC colonies and establish colony characteristics database.
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-019-53719-9