Introduction
My name is I.J. Nzeama, and I am a final-year Biomedical Engineering student at Nottingham Trent University. With support from the RS Student Fund, I undertook a challenging but deeply rewarding research project: the development of a three-dimensional (3D) breast cancer organoid model designed to better reflect tumour behaviour in the human body.
Breast cancer is the most diagnosed cancer globally and a leading cause of mortality among women. Yet, many of the tools we use in the lab to study it, particularly 2D cell cultures, unfortunately fail to replicate the complexity of real tumours. This project aimed to bridge that gap.
Project Aim
The objective of this research was to design a physiologically relevant 3D in vitro breast cancer model that mimics the tumour microenvironment. Such a model could be used to improve drug screening accuracy and enhance our understanding of tumour–stromal interactions.
Success was measured by the viability, structure, and behaviour of the co-cultured spheroids over time, and how well they responded to treatment in controlled conditions.
Methodology Overview
To construct this organoid system, I co-cultured MCF-7 breast cancer cells with fibroblasts, embedding them within an extracellular matrix (ECM) to mimic the tumour environment. I tested various centrifugation speeds and cell seeding densities to optimise spheroid formation.
Two key protocols emerged:
- High speed (13,000 rpm) with 40,000 cells: Promoted early spheroid compactness and initial viability.
- Low speed (1,000 rpm) with 60,000 cells: Supported longer-term survival and growth.
To assess the model, I used:
- MTT assays to evaluate metabolic activity
- Live/Dead staining to monitor cell viability over time
- Immunocytochemistry (ICC) to assess cellular arrangement within spheroids
Development Process
Initial trials highlighted several challenges in forming consistent, robust spheroids. Some conditions caused weak aggregation or rapid disintegration. Iterative refinement led to embedding spheroids within ECM for added structure and physiological relevance.
MTT testing on Days 1, 3, and 7 revealed that:
- High-speed conditions showed strong early activity but plateaued over time.
- Low-speed conditions demonstrated growing activity, suggesting better nutrient diffusion and cell interaction.
- Live/Dead staining across Days 7 and 21 showed minimal cell death early on, but increased core necrosis and disrupted architecture by Day 21, likely due to static culture limitations.
- ICC imaging on Day 1 revealed intriguing cellular organisation:
- Fibroblasts tended to localise centrally.
- MCF-7 cells formed a ring around the periphery, mirroring in vivo spatial patterns
Challenges and Solutions
Structural Integrity
Early spheroids lacked cohesion. ECM embedding improved spheroid stability and shape retention.
Viability Over Time
By Day 21, central necrosis increased—highlighting the need for improved nutrient and waste exchange systems (e.g. perfusion culture).
Spatial Complexity
Unexpectedly, ICC revealed a degree of cell self-organisation. This finding offers promise for studying tumour-stromal dynamics in future iterations.
Impact of the RS Student Fund
The RS Student Fund made this research possible. The funding allowed access to:
- Centrifuges and fluorescence microscopes
- High-quality staining reagents
- ECM products for 3D culture
- Specialist laboratory spaces
This support elevated the scope and professionalism of my research, giving me access to real-world tools and laboratory infrastructure typically reserved for industry or postgraduate researchers.
Outcomes and Future Applications
This project successfully established a reproducible method for generating co-culture breast cancer spheroids with features that reflect tumour complexity. The model:
- Demonstrates time-dependent metabolic and viability shifts
- Exhibits differential drug responses
- Provides insight into tumour–stromal architecture
In the future, this platform could support:
- Drug screening in early-stage research
- Incorporation of immune cells or patient-derived cells, Personalised treatment testing in oncology
Reflections and Advice for Future Students
This project taught me that research is a design process, an act of problem-solving under uncertainty. There were many failed attempts, but each one offered insight. If you’re considering your own project, my advice is:
- Be patient with setbacks as they often reveal what matters most.
- Don’t be afraid to trial unconventional methods.
- Seek out funding and support early—it can transform what you’re able to do.
- The RS Student Fund doesn’t just enable projects; it empowers students to think and act like engineers and innovators.
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