3D Tumor Model for Breast Cancer
Alfa Cytology offers advanced solutions for breast cancer cell models, addressing the challenges in constructing 3D breast cancer models. Our 3D models better represent the tumor physiology compared to traditional 2D culture systems. With our extensive cancer model database, our team can assist in selecting and designing studies for optimal outcomes using 3D assays.
Overview of 3D Tumor Models for Breast Cancer
In the past decade, there has been remarkable growth in the development of 3D models for various types of cancer, including diverse 3D breast cancer models. Porous gelatin microspheres were utilized to create breast cancer models. The beads were seeded with dermal fibroblasts, HUVECs, and ultimately MCF-10 or MCF-7 cells. Angiogenesis was assessed based on the branching vessel diameter.
Fig.1 3D printing of complex structures for in vitro tumor modeling. (Pasquier E, et al., 2023)
Our Services
Alfa Cytology offers a range of 3D models specifically designed for studying tumor physiology and conducting drug screening in breast cancer research. These models encompass various types, such as spheroids, organoids, breast cancer on a chip, and 3D bioprinting of tissues. We provide spheroids, organotypic cultures, and organoid models to facilitate the exploration of breast cancer and the development of new therapeutics.
Spheroids Models
Cell aggregates known as spheroids can be cultured in suspension, either on low-adhesion plastic plates or inert substrates like agarose, with continuous agitation. In the system, cancer cells are deposited on an ultralow fixation plate, which effectively prevents adhesion and facilitates suspension-based cell growth.
Breast cancer cells are seeded onto a biocompatible scaffold that mimics the extracellular matrix. This scaffold provides a three-dimensional structure that promotes cell adhesion and growth. The scaffold model is used to study tumor development, cellular interactions, and drug responses, offering a controlled environment for breast cancer research.
Combine human primary breast cancer organoids with adipose tissue to generate an immunocompetent breast cancer chip model. This tumor chip can serve as a standardized, scalable, and exploratory platform for assessing the efficacy of drugs.
The iPSCs undergo a reprogramming process, followed by directed differentiation. Subsequently, these cells are seeded into a specialized culture medium, along with an extracellular matrix, to initiate organoid culture. They are then seeded on Matrigel.
Construction of 3D Breast Cancer Models
Selecting the Right Cell Line
Considering the diverse phenotypic manifestations of breast cancer, we offer representative cell lines for cell culture, such as T47D and MDA-MB-231 cell lines.
Formation of uniform cell spheroids
After reaching the desired degree of cell confluence, single-layer cells are detached from the surface of cell culture using cell dissociation reagents, resulting in the formation of a single-cell suspension.
Optimize growth conditions for specific cell types
To facilitate the formation of spheroids, various optimizations are implemented in terms of culture media and cell seeding conditions, taking into account the varying abilities of different cell types to form spherical structures.
Cell Health Analysis
Prior to utilizing spheroids for downstream assays, a Cell Health Analysis is conducted, which involves assessing the size, compactness, cellular health, and viability of the spheroids.
Dose Response Assessment
Once the assessment of cellular health and spheroid activity is completed, the viable spheroids can be utilized for various downstream assays and corresponding research studies.
Case Study
T47D Breast Cancer Spheroid Model for Drug Screening
- Model Introduction
The T47D cell line-based spheroid model serves as a physiologically relevant in vitro platform for evaluating drug efficacy, screening therapeutic agents, and studying tumor biology in estrogen receptor-positive (ER+) breast cancer. It is particularly valuable for high-throughput drug susceptibility testing, cytotoxicity assessment, and personalized treatment strategy exploration.
- Model Information
- Model: T47D 3D Spheroid Model
- Cell Line: T47D Human Breast Cancer Cells
- Cancer Type: Invasive Ductal Carcinoma (IDC)
- Culture Medium: RPMI-1640 with 20% FBS
- Model Construction
The three-dimensional spheroid model was established by seeding T47D human breast cancer cells onto ultra-low attachment microplates, including:
- Culture: T47D Cells were maintained and expanded as 2D monolayers in RPMI-1640 medium supplemented with 20% FBS using standard tissue culture-treated flasks.
- Generate: To initiate spheroid formation, a single-cell suspension was prepared using a cell dissociation reagent and seeded onto ultra-low attachment microplates.
- Optimize: Through optimized culture conditions with a critical seeding density of ≤8,000 cells per well in 96-well plates.The T47D cell line demonstrated excellent spontaneous self-assembly capability under the defined conditions.
Fig. 2 Bright-field image of T47D cell aggregates and spheroids on day 2. (Source: Alfa Cytology)
- Characterize: Under optimized conditions, T47D cells consistently self-assembled into uniform, compact spheroids with an approximate diameter of 600 µm within 3-4 days. Spheroid size, roundness, and overall morphology were monitored using live-cell imaging systems.
- Measure: After assessing cell health and spheroid activity, cell proliferation assays were performed using healthy spheroids to validate the T47D spheroids model.
- Model Data
- Consistent Spheroid Formation: The model demonstrates reliable and reproducible self-assembly capability, forming uniform three-dimensional structures with well-defined morphology suitable for quantitative drug screening.
- Controlled Proliferation Kinetics: Analysis of four-day-old T47D spheroids treated with 100 nM drug A for 72 hours using Click-iT EdU assay revealed distinct proliferation patterns, confirming the model's utility for therapeutic efficacy assessment.
Fig. 3 Characterization and determination of T47D sphere cells. (Source: Alfa Cytology)
Contact Us
Alfa Cytology is dedicated to the development of breast cancer therapies and possesses extensive expertise in constructing 3D tumor models. Through collaboration with Alfa Cytology, our 3D cell culture models can support your endeavors in oncology and immuno-oncology drug development. For further information regarding our services, please reach out to our team for expert advice.
Reference
- Pasquier E, et al. Polysaccharides and Structural Proteins as Components in Three-Dimensional Scaffolds for Breast Cancer Tissue Models: A Review. Bioengineering (Basel). 2023, 10(6):682.