Abstract
Given the difficulties inherent in maintaining human pluripotent stem cells (hPSCs) in a healthy state, hPSCs should be routinely characterized using several established standard criteria during expansion for research or therapeutic purposes. hPSC colony morphology is typically considered an important criterion, but it is not evaluated quantitatively. Thus, we designed an unbiased method to evaluate hPSC colony morphology. This method involves a combination of automated non-labelled live-cell imaging and the implementation of morphological colony analysis algorithms with multiple parameters. To validate the utility of the quantitative evaluation method, a parent cell line exhibiting typical embryonic stem cell (ESC)-like morphology and an aberrant hPSC subclone demonstrating unusual colony morphology were used as models. According to statistical colony classification based on morphological parameters, colonies containing readily discernible areas of differentiation constituted a major classification cluster and were distinguishable from typical ESC-like colonies; similar results were obtained via classification based on global gene expression profiles. Thus, the morphological features of hPSC colonies are closely associated with cellular characteristics. Our quantitative evaluation method provides a biological definition of ‘hPSC colony morphology’, permits the non-invasive monitoring of hPSC conditions and is particularly useful for detecting variations in hPSC heterogeneity.
Highlights
(high in volume parameters), with a boundary that is not round, and a very irregular morphology
201B7-1A subclone that gained an additional copy of chromosome 12 after prolonged culture and its healthy parent 201B7 line, which was the first human induced pluripotent stem cells (hiPSCs) line reported by Yamanaka’s group[2] (Supplementary Tables S1–3 and Supplementary Fig. S1)
HPSC colony morphology is a practical and key criterion to assess the quality of Human pluripotent stem cells (hPSCs) maintained under laboratory conditions
Summary
(high in volume parameters), with a boundary that is not round (low in shape parameters), and a very irregular morphology (low in frequency). The technical combination of ‘massive and stable non-biased images of cells’ and ‘multi-parametric analysis’ permits the analysis of cellular morphological features and replaces analyses dependent on conventional cell biological features such as marker profiles or gene expression determined via invasive methods. We propose a non-invasive image-based evaluation method for detecting partially differentiated colony morphology in heterogeneous colony populations via live image analysis. A parent cell line exhibiting typical embryonic stem cell (ESC)-like morphology and an aberrant hPSC subclone (#12 trisomy) exhibiting partially differentiated colony morphology were used as models to validate the utility of our quantitative evaluation method. Colony morphology classification based on the statistical analysis of non-labelled live-cell images with unbiased morphological parameters was compared with classification based on global gene expression profiles of individual colonies. Our quantitative morphological evaluation method facilitates the non-invasive analysis of hiPSC conditions and demonstrates particular utility in monitoring variations in hPSC heterogeneity
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
