This invention presents an innovative method for forming cell spheroids using three-dimensional cryogel polymer matrices composed of polyethylene glycol diacrylate (PEGDA), gelatin methacrylate (GELMA), and/or alginate methacrylate (ALMA). The 3D cryogel matrix technology allows for the creation of cell spheroids that closely mimic the natural environment of tissues and tumors. This method is both cost-effective and efficient, eliminating the need for additional growth factors. It provides a stable, reproducible, and physiologically relevant substrate for in vitro cell culture studies, making it a valuable tool for applications in biomedical research, pharmaceutical development, tissue engineering, and cancer research.
Figure (1) Confocal fluorescence images showing morphology of cells stained with Phalloidin and DAPI at day 7
Traditional 2D cell culture methods fail to accurately mimic the native morphology and environment of tissues or tumors. This leads to limitations in understanding cell behavior, disease mechanisms, drug actions, and responsiveness to stimuli, especially in cancer research where the 3D architecture of tumors significantly impacts nutrient and oxygen availability. Existing 3D cell culture techniques also suffer from issues like poor reproducibility, instability, complexity, difficulty in scaling, and a need for more physiologically relevant substrates. There is a need for a low-cost, reproducible, and stable 3D matrix that can facilitate the formation of cellular spheroids without external growth factors, providing a better representation of in vivo conditions for drug screening and disease studies.
- Innovative Cryogel Composition: A cryogel matrix formulation using a specific combination of PEGDA, GELMA, and ALMA polymers enhances the mechanical properties and biocompatibility of the matrix, facilitating better cell adhesion and proliferation.
- Temperature-Controlled Cryopolymerization: A precise temperature-controlled cryopolymerization process allows for the formation of highly porous and interconnected cryogel structures. This feature supports efficient nutrient and oxygen diffusion, closely mimicking the in vivo environment.
- Scalable Matrices and improved Reproducibility: The cryogel matrices are designed to be easily scalable and customizable to fit various experimental needs. High reproducibility is achieved through stringent control of the polymerization and cross-linking processes.
- Enhanced Spheroid Stability: Advanced cross-linking techniques using ammonium persulphate (APS) and tetramethylethylenediamine (TEMED) significantly enhanced the stability and longevity of the formed spheroids.
- Elimination of Growth Factors: By optimizing the matrix composition and structure, we eliminated the need for additional growth factors, reducing both the cost and complexity of cell culture experiments. This innovation simplifies the setup and makes the technology more accessible to a broader range of laboratories.
Successful fabrication and testing of the cryogel matrices and the successful development of spheroids. This includes specific material concentrations, the freezing temperature (-20°C) and duration (18 hours) for cryopolymerization, and the range of cell densities (500 to 10 6 ^ cells per cryogel) used to seed the matrices. The results, evidenced by morphological studies, cell proliferation, and comparative drug resistance analyses, demonstrate the functionality and efficacy of this specific prototype formulation and method in generating and utilizing the claimed 3D cellular spheroids.
This technology enables efficient formation of cell spheroids using 3D cryogel matrices made from PEGDA, GELMA, and/or ALMA. It closely mimics native tissue environments, is cost-effective, and eliminates the need for external growth factors. We have successfully developed in vitro breast cancer models using this platform and are now testing it for our tissue models. It offers a stable, reproducible system for applications in cancer research, tissue engineering, and drug screening.
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The development of a more accurate and reliable 3D cell culture system can significantly advance research in cell biology, disease mechanisms, and drug development. This can lead to better understanding and treatment of diseases, more effective drugs, and potential breakthroughs in tissue engineering and regenerative medicine.
Biomedical Research, Pharmaceutical Development, Biotechnology, Tissue Engineering, Cancer Research, Regenerative medicine, Cosmetics Industry.
Geography of IP
Type of IP
201921011783
520387