Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2022 Apr 22;12:818437. doi: 10.3389/fonc.2022.818437

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

Copyright © 2022 Lotsberg, Røsland, Rayford, Dyrstad, Ekanger, Lu, Frantz, Stuhr, Ditzel, Thiery, Akslen, Lorens and Engelsen

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

PMC Copyright notice

Monitoring cell aggregation and 3D spheroid formation ability in real time using the IncuCyte live cell imaging system. (A) The lung adenocarcinoma cell line HCC827 and erlotinib-resistant sub-clones of this cell line (ER3, ER10) were cultured as homotypic spheroids (upper panel) and in combination with lung fibroblast cell lines SV80 (middle panel) or MRC-5 (lower panel). Images were obtained with the IncuCyte live cell imaging system 24 h after seeding in the ultra-low attachment (3D) 96-well plates. The IncuCyte confluence mask (yellow) was generated for the quantification shown in (C, D). (B) The NSCLC cell line H1975 and rociletinib-resistant sub-clones (COR1-1 and COR10-1) were cultured as homotypic spheroids (upper panel) or in combination with lung fibroblast cell lines SV80 (middle panel) or MRC-5 (lower panel). Images were obtained with the IncuCyte live cell imaging system 24 h after seeding in the ultra-low attachment (3D) 96-well plates. Spheroid formation efficiency was measured by quantification of the object confluence measured by the IncuCyte Zoom microscope and software-generated confluence mask (yellow) in HCC827, ER3, or ER10 cell monocultured spheroids or as heterotypic co-culture spheroids together with (C) SV80 or (D) MRC-5. Object confluence over the 24 h time-course and the values for the 24 h endpoint is given. Spheroid formation assays were repeated at least three times, and representative results from one experiment with n = 6-10 technical replicates are presented as mean +/- SD. One-way ANOVA followed by Tukey’s multiple comparisons test was performed to calculate statistical differences in object confluence at 24 h. ****P ≤ 0.0001. ns, not significant.