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Generation of MCF-7 Spheroids in Polyethylene Glycol-Dextran Droplets for Cancer Niche Studies Using Aqueous
Karolina Chairez-Cantu1, Mirna González-González1,2, Marco Rito-Palomares1,2
1Tecnologico de Monterrey, Escuela de Medicina y Ciencias de la Salud, Monterrey, Nuevo Leon, Mexico.
Biotechnology Journal
|April 14, 2025
Summary
This study developed 3D cell culture models using polyethylene glycol (PEG)-dextran (DEX) aqueous two-phase systems (ATPS) for cancer research. The optimized PEG-DEX ATPS demonstrated enhanced cell viability and tumor-specific traits, suitable for drug screening.
Area of Science:
- Biotechnology
- Cell Biology
- Cancer Research
Background:
- Cell-based aqueous two-phase systems (ATPS) enable 3D cell culture by encapsulating cells in polyethylene glycol (PEG)-dextran (DEX) droplets.
- This technology facilitates controlled cell patterning and the creation of specific cell microenvironments.
Purpose of the Study:
- To encapsulate MCF-7 cancer cells in PEG-DEX ATPS droplets.
- To evaluate different PEG-DEX compositions, construction strategies (A, B, C), and cell densities.
- To assess spheroid formation, droplet characteristics, and cell viability over 24 hours and 4 days.
Main Methods:
- Utilized 96-well plates for ATPS droplet formation with varying PEG-DEX compositions and construction strategies.
- Encapsulated MCF-7 cells at densities of 5000 and 10,000 cells/µL.
- Analyzed droplet size, circularity, spheroid formation, and cell viability over time.
Main Results:
- Droplet size varied by construction strategy, with Strategy C yielding smaller droplets (1.05 ± 0.33 mm) and Strategy B larger ones (1.46 ± 0.49 mm). Strategy A showed higher circularity.
- MCF-7 spheroids formed within 24 hours, with diameters ranging from 0.10 to 0.65 mm, exhibiting higher circularity at lower cell densities.
- The PEG 35,000-DEX 500,000 system demonstrated higher cell viability for 4 days, displaying tumor-specific traits compared to 2D cultures.
Conclusions:
- Demonstrated practical strategies for constructing 3D cancer models using PEG-DEX ATPS with standard laboratory equipment.
- The developed 3D models show promise for future drug screening studies by mimicking the cancer niche.
- Optimized PEG-DEX compositions and construction methods can enhance cell viability and tumor-specific characteristics in 3D cultures.
