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High-throughput formation and image-based analysis of basal-in mammary organoids in 384-well plates.
- Source :
-
Scientific reports [Sci Rep] 2022 Jan 10; Vol. 12 (1), pp. 317. Date of Electronic Publication: 2022 Jan 10. - Publication Year :
- 2022
-
Abstract
- This manuscript describes a new method for forming basal-in MCF10A organoids using commercial 384-well ultra-low attachment (ULA) microplates and the development of associated live-cell imaging and automated analysis protocols. The use of a commercial 384-well ULA platform makes this method more broadly accessible than previously reported hanging drop systems and enables in-incubator automated imaging. Therefore, time points can be captured on a more frequent basis to improve tracking of early organoid formation and growth. However, one major challenge of live-cell imaging in multi-well plates is the rapid accumulation of large numbers of images. In this paper, an automated MATLAB script to handle the increased image load is developed. This analysis protocol utilizes morphological image processing to identify cellular structures within each image and quantify their circularity and size. Using this script, time-lapse images of aggregating and non-aggregating culture conditions are analyzed to profile early changes in size and circularity. Moreover, this high-throughput platform is applied to widely screen concentration combinations of Matrigel and epidermal growth factor (EGF) or heparin-binding EGF-like growth factor (HB-EGF) for their impact on organoid formation. These results can serve as a practical resource, guiding future research with basal-in MCF10A organoids.<br /> (© 2022. The Author(s).)
- Subjects :
- Algorithms
Cell Line
Collagen pharmacology
Drug Combinations
Epidermal Growth Factor pharmacology
Female
Heparin-binding EGF-like Growth Factor pharmacology
Humans
Laminin pharmacology
Mammary Glands, Human cytology
Mammary Glands, Human drug effects
Proteoglycans pharmacology
Time Factors
Cell Culture Techniques, Three Dimensional instrumentation
Cell Proliferation drug effects
High-Throughput Screening Assays
Image Processing, Computer-Assisted
Mammary Glands, Human physiology
Microscopy, Fluorescence
Organoids
Time-Lapse Imaging
Subjects
Details
- Language :
- English
- ISSN :
- 2045-2322
- Volume :
- 12
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Scientific reports
- Publication Type :
- Academic Journal
- Accession number :
- 35013350
- Full Text :
- https://doi.org/10.1038/s41598-021-03739-1