3D Tumor Spheroid Culture
Making Your Cancer Research More Physiologically Relevant
The human body is a 3D system, yet most cell culture research uses 2D monolayers. 2D cultures do not accurately reflect cell attachment, cytoskeletal structure, or the in vivo behavior of cells. 3D spheroids provide a more physiologically representative model than traditional culture systems and can be used in many applications, including cancer research, drug screening, and stem cell research.
3D tumor spheroids, also known as cancer spheroids, have been shown to mimic the following parameters of in vivo tumors:
- Study tumorigenesis, spheroid morphology, self-organization, cytoskeletal structure, signalling, and cancer progression.
- Reproduce gradients of oxygen and nutrient availability throughout the tumor, and study roles of outer vs. inner tumor cells.
- Model 3D cell-cell and cell-ECM interactions, and co-culture to identify roles of stromal cells and cancer-associated fibroblasts.
- Study epithelial-mesencyhmal transitions, cell migration, and survival of cancer in secondary sites.
- Predict drug response, drug kinetics, cytotoxicity, and diffusion for solid tumor-based studies of drug discovery and toxicity.
- Model cytotoxic effects of T cells or NK cells using a 3D tumor model.
Reproducible 3D Spheroid Cultures Using ´¡²µ²µ°ù±ð°Â±ð±ô±ôâ„¢ Plates
´¡²µ²µ°ù±ð°Â±ð±ô±ôâ„¢ microwell plates make it easy to generate hundreds of uniformly-sized 3D spheroids. Simply add a single-cell suspension to the wells containing microwells and centrifuge to distribute the cells evenly in the microwells. Incubate overnight and assess the spheroids after 24 - 48 hours. Then culture in the microwells, or harvest and perform downstream assays.
´¡²µ²µ°ù±ð°Â±ð±ô±ôâ„¢ helps ensure uniformity of spheroids and consistency from experiment to experiment.
´¡²µ²µ°ù±ð°Â±ð±ô±ôâ„¢ plates with microwells provide an easy way to generate large numbers of multicellular spheroids. Unlike traditional spheroid-formation methods, spheroids generated in ´¡²µ²µ°ù±ð°Â±ð±ô±ôâ„¢ are highly uniform in size and shape, and the size can be easily controlled by modifying the input cell seeding density. With added size-control and uniformity, AggreWell™ improves reproducibility of your 3D research.
Why use ´¡²µ²µ°ù±ð°Â±ð±ô±ôâ„¢?
- Generate large numbers of spheroids from a single pipetting step compared to at least 8 pipetting steps with a 96-or 384-well ULA or hanging drop plate. Fewer processing steps means fewer opportunities for error.
- Spheroids generated in ´¡²µ²µ°ù±ð°Â±ð±ô±ôâ„¢ are highly uniform in size and shape, meaning that your starting population is consistent within and between experiments. Get true technical replicates, without compounding population or processing effects.
- Large numbers of spheroids facilitate larger, more comprehensive studies with statistically significant sample sizes. A statistically significant sample size increases the scientific accuracy and reproducibility of your experimental results.
´¡²µ²µ°ù±ð°Â±ð±ô±ôâ„¢ helps you ensure the clonality of your sphere-forming assay.
In the sphere-forming assay, spheres are generated from a single cell, often referred to as a Tumor-Initiating Cell (TIC) or a Cancer Stem-like Cell (CSC), which is capable of self-renewal and proliferation. The sphere-forming assay is a common method used to determine the frequency of the TICs in a given cancer cell population. However, achieving clonality requires limiting dilution, which can be difficult and laborious. By using ´¡²µ²µ°ù±ð°Â±ð±ô±ôâ„¢ with microwells, clonal spheres are easy to achieve without the need for limiting dilution.
Why use ´¡²µ²µ°ù±ð°Â±ð±ô±ôâ„¢?
- With 1,200 microwells in each well (´¡²µ²µ°ù±ð°Â±ð±ô±ôâ„¢400 24-well), there are 1,200 opportunities for each cell to settle into a microwell and be physically sequestered away from other cells, preventing migration and aggregation.
- Get clonal results by seeding at limiting dilution densities in a single pipetting step.
- At limiting dilution cell density and the ability to visually confirm single cells in microwells, the accuracy of your clonal sphere-forming assay can be verified.
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References
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