Live-cell imaging of subcellular structures for quantitative evaluation of pluripotent stem cells

Abstract

Pluripotent stem cells (PSCs) have various degrees of pluripotency, which necessitates selection of PSCs with high pluripotency before their application to regenerative medicine. However, the quality control processes for PSCs are costly and time-consuming, and it is essential to develop inexpensive and less laborious selection methods for translation of PSCs into clinical applications. Here we developed an imaging system, termed Phase Distribution (PD) imaging system, which visualizes subcellular structures quantitatively in unstained and unlabeled cells. The PD image and its derived PD index reflected the mitochondrial content, enabling quantitative evaluation of the degrees of somatic cell reprogramming and PSC differentiation. Moreover, the PD index allowed unbiased grouping of PSC colonies into those with high or low pluripotency without the aid of invasive methods. Finally, the PD imaging system produced three-dimensional images of PSC colonies, providing further criteria to evaluate pluripotency of PSCs. Thus, the PD imaging system may be utilized for screening of live PSCs with potentially high pluripotency prior to more rigorous quality control processes.

Mechanisms of the Metabolic Shift during Somatic Cell Reprogramming

“…Nevertheless, these studies point to an essential, and possibly regulatory, role for mitochondrial changes during reprogramming. Indeed, the morphological differences between partially and fully reprogrammed can be quantifiable in live cells by a modified retardation modulated-differential interference contrast (RM-DIC) microscope and can be used to accurately predict the pluripotency of derived iPSCs [86]. All of these studies indicate a close relationship between remodeling of mitochondria and acquisition of pluripotency, and further study is warranted for understanding more precise mechanisms of mitochondrial remodeling and its role in reprogramming.…”

Video Bioinformatics Analysis of Human Pluripotent Stem Cell Morphology, Quality, and Cellular Dynamics

“…10 Time-lapse imaging has also been used to study cells that are undergoing reprogramming to better understand the dynamics of the reprogramming process. 27,28 Because of their small cytoplasmic to nuclear ratio, the number of cells in hESC colonies is difficult to determine, but cell counting in colonies has been successfully done using label-free time-lapse images. 8 New PSC culture media are continually being developed and tested with hPSC, as well as with embryonic stem cells from other species, 29,30 and StemCellQC could be used to help evaluate the performance of such media.…”

Decoding Activation of ILC2 using Time-Dependent Cell-State Selection

“…While we focused on the activation process of ILC2 in this study, many LCI techniques have been developed to trace various fate determination processes, including immune response 16 , cell differentiation 42,43 , cell death 2 , and stimulus response 10,44 . Using these LCI techniques, the TDCSS technique will help elucidating contexts of these fate determination processes as well.…”