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Invasion Analysis and RNA Analysis : Prepare gels for 240 wells for approximately ¥10,000.
― From live imaging to full transcriptome profiling ―
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Seeing is believing. Capturing experimental results as images not only strengthens the impact of your research but also enables direct comparison of changes over time. In addition, combining image analysis with microarrays generates quantitative data that reinforce scientific evidence.
Here, we highlight research using real-time invasion imaging of glioblastoma spheroids in atelocollagen gel paired with transcriptome analysis.
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Four Steps to Prepare for Cancer Cell Invasion and Transcriptome Analysis
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The panels above depict a method for evaluating cancer cell invasion divided into four clear steps:
(1) Generation of spheroids, (2) Embedding of spheroids in atelocollagen gel, (3) Capturing live-cell images, and (4) Extraction of total RNA for transcriptome analysis.
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Cancer Cell Lines Suitable for Spheroid Formation
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The glioblastoma multiforme cell line (SF126) formed compact spherical spheroids in ultra-low attachment plates (left photo). In contrast, some cell lines such as pancreatic cancer cells (MIA PaCa-2), produced dispersed aggregates (right photo), making them less suitable for evaluating cancer cell invasion from spheroids.
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Quantifying the Dispersion of Invaded Cells in Atelocollagen Gel Over Time
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When SF126 spheroids were embedded in atelocollagen gel and cultured, invasive cancer cells invading from the spheroid were observed.
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This behavior contrasts sharply with the control cells cultured in ultra-low attachment plates.
The system can also be used to test the inhibitory effects of compounds on cancer cell invasion by adding compounds directly to the same culture system.
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Collecting Total RNA After Evaluating the Dispersion of Invaded Cells
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For comprehensive gene expression analysis of post-invasion cells, atelocollagen gels from ultra-low attachment plates can be transferred to tubes, and total RNA can then be extracted using a standard commercial RNA extraction kit.
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Transcriptome Analysis and RT-qPCR Validation
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Comprehensive gene expression profiling was performed using the extracted total RNA. Gene Ontology analysis revealed reduced expression of cell cycle–related genes and increased expression of cell migration–related genes. RT-qPCR validation further confirmed these changes in gene expression.
Source：STAR Protoc. 2025 Jun 20;6(2):103839. Created by modifying graphical abstract, figure 1, 5, 6 and 7. ©Fujita M., et al. 2025.
Licensed under CC BY 4.0 (https://creativecommons.org/licenses/by/4.0/).
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Atelocollagen Solution Used in the Aforementioned Paper
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This ready-to-use atelocollagen solution gels simply upon warming, enabling easy and stable 3D culture for any user. One bottle is sufficient for testing 240 wells in a 96-well format.
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Exhibition Information
We plan to participate in the annual meeting of the Cancer Association in September 2025. Please visit us at the venue to see our Atelocollagen products.
84th Annual Meeting of the Japanese Cancer Association
Dates: September 25 (Thurs.)–27 (Sat.) 2025
Venue: B1F Koryu Hall, Ishikawa Ongakudo
Booth: K-19
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