An integrated pipeline for high-throughput screening and profiling of spheroids using simple live image analysis of frame to frame variations.

Details

Serval ID
serval:BIB_9E1ED23C83DC
Type
Article: article from journal or magazin.
Collection
Publications
Title
An integrated pipeline for high-throughput screening and profiling of spheroids using simple live image analysis of frame to frame variations.
Journal
Methods
Author(s)
Alsehli H., Mosis F., Thompson C., Hamrud E., Wiseman E., Gentleman E., Danovi D.
ISSN
1095-9130 (Electronic)
ISSN-L
1046-2023
Publication state
Published
Issued date
06/2021
Peer-reviewed
Oui
Volume
190
Pages
33-43
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: ppublish
Abstract
High-throughput imaging methods can be applied to relevant cell culture models, fostering their use in research and translational applications. Improvements in microscopy, computational capabilities and data analysis have enabled high-throughput, high-content approaches from endpoint 2D microscopy images. Nonetheless, trade-offs in acquisition, computation and storage between content and throughput remain, in particular when cells and cell structures are imaged in 3D. Moreover, live 3D phase contrast microscopy images are not often amenable to analysis because of the high level of background noise. Cultures of Human induced pluripotent stem cells (hiPSC) offer unprecedented scope to profile and screen conditions affecting cell fate decisions, self-organisation and early embryonic development. However, quantifying changes in the morphology or function of cell structures derived from hiPSCs over time presents significant challenges. Here, we report a novel method based on the analysis of live phase contrast microscopy images of hiPSC spheroids. We compare self-renewing versus differentiating media conditions, which give rise to spheroids with distinct morphologies; round versus branched, respectively. These cell structures are segmented from 2D projections and analysed based on frame-to-frame variations. Importantly, a tailored convolutional neural network is trained and applied to predict culture conditions from time-frame images. We compare our results with more classic and involved endpoint 3D confocal microscopy and propose that such approaches can complement spheroid-based assays developed for the purpose of screening and profiling. This workflow can be realistically implemented in laboratories using imaging-based high-throughput methods for regenerative medicine and drug discovery.
Keywords
Cell Culture Techniques, High-Throughput Screening Assays, Humans, Induced Pluripotent Stem Cells, Microscopy, Confocal, Spheroids, Cellular, 3D, Cell phenotyping, High content imaging, High throughput imaging, Spheroids, Stem cells
Pubmed
Web of science
Open Access
Yes
Create date
12/01/2024 11:14
Last modification date
13/01/2024 8:10
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