X-ray microtomography as a new approach for imaging and analysis of tumor spheroids
Hanieh Karimi1, Bartosz Leszczyński1, Tomasz Kołodziej2
1Department of Medical Physics, M. Smoluchowski Institute of Physics, Faculty of Physics, Astronomy and Applied Computer Science, Jagiellonian University, Kraków, Poland.
Abstract:
Three-dimensional (3D) spheroids mimic important properties of tumors and may soon become a reasonable substitute for animal models and human tissue, eliminating numerous problems related to in vivo and ex vivo experiments and pre-clinical drug trials. Currently, various imaging methods including X-ray microtomography (micro-CT), exist but their spatial resolution is limited. Here, we visualized and provided a morphological analysis of spheroid cell cultures using micro-CT and compared it to that of confocal microscopy. An approach is proposed that can potentially open new diagnostic opportunities to determine the morphology of cancer cells cultured in 3D structures instead of using actual tumors. Spheroids were formed from human melanoma cell lines WM266-4 and WM115 seeded at different cell densities using the hanging drop method. Micro-CT analysis of spheroid showed that spheroid size and shape differed depending on the cell line, initial cell number, and duration of culture. The melanoma cell lines used in this study can successfully be cultured as 3D spheroids and used to substitute human and animal models in pre-clinical studies. The micro-CT allows for high-resolution visualization of the spheroids structure.
Insights
Three-dimensional (3D) spheroids offer a promising alternative to animal models for cancer research. Micro-X-ray computed tomography (micro-CT) provides high-resolution visualization of these 3D cell cultures, aiding in morphological analysis.
Area of Science:
- Biomedical Engineering
- Cancer Research
- Cell Biology
Background:
- Three-dimensional (3D) spheroids closely mimic tumor properties, presenting a potential replacement for animal models and human tissues in pre-clinical studies.
- Current imaging techniques for spheroids, such as micro-X-ray computed tomography (micro-CT), have limitations in spatial resolution.
- Developing advanced imaging methods is crucial for accurate morphological analysis of 3D cell cultures.
Purpose of the Study:
- To visualize and morphologically analyze spheroid cell cultures using micro-CT.
- To compare the efficacy of micro-CT with confocal microscopy for spheroid imaging.
- To assess the potential of 3D spheroids as substitutes for traditional pre-clinical models.
Main Methods:
- Human melanoma cell lines (WM266-4 and WM115) were cultured into 3D spheroids using the hanging drop method at varying cell densities.
- Micro-X-ray computed tomography (micro-CT) was employed for high-resolution imaging and morphological analysis of the generated spheroids.
- Confocal microscopy was used as a comparative imaging technique.
Main Results:
- Micro-CT analysis revealed that spheroid size and shape varied based on cell line, initial cell number, and culture duration.
- The study demonstrated successful culturing of melanoma cell lines as 3D spheroids.
- Micro-CT enabled high-resolution visualization of the internal structure of the spheroids.
Conclusions:
- Human melanoma cell lines can be effectively cultured as 3D spheroids for pre-clinical applications.
- Micro-CT offers a valuable tool for high-resolution morphological assessment of 3D spheroid structures.
- This approach holds potential for new diagnostic opportunities in analyzing cancer cell morphology within 3D structures.
