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This technology has enabled cell patterning in a controlled\u2010defined shape giving rise to specific cell microenvironments. The present work aims to encapsulate MCF\u20107 cancer cells in ATPS droplets evaluating four different compositions of PEG\u2010DEX, construction Strategies A (droplet added), B (immersed), or C (covered), and two cell densities (5000 and 10,000 cells\/\u00b5L) during a 24 h incubation time using 96\u2010well plates. The smallest cell\u2010containing DEX droplets showed a mid\u2010range value of 1.05\u00a0\u00b1\u00a00.33\u00a0mm using Strategy C whereas the largest of 1.46\u00a0\u00b1\u00a00.49\u00a0mm in Strategy B. However, Strategy A was associated with higher rates of circularity. After 24 h, MCF\u20107 spheroids were formed in all PEG\u2010DEX systems showing a diameter size ranging from 0.10 to 0.65\u00a0mm. Their circularity was higher in low cell densities. Higher rates of cell viability were obtained in PEG 35,000\u2013DEX 500,000 systems for a period of 4 days showing tumor\u2010specific traits compared to 2D control. This work demonstrates practical strategies for 3D model construction for cancer niche for future drug screening studies using basic laboratory equipment.<\/jats:p>","DOI":"10.1002\/biot.70014","type":"journal-article","created":{"date-parts":[[2025,4,14]],"date-time":"2025-04-14T05:01:16Z","timestamp":1744606876000},"source":"Crossref","is-referenced-by-count":1,"title":["Generation of MCF\u20107 Spheroids in Polyethylene Glycol\u2010Dextran Droplets for Cancer Niche Studies Using Aqueous Two\u2010Phase System\u20103D Platforms"],"prefix":"10.1002","volume":"20","author":[{"given":"Karolina","family":"Chairez\u2010Cantu","sequence":"first","affiliation":[{"name":"Tecnologico de Monterrey Escuela de Medicina y Ciencias de la Salud  Monterrey Nuevo Leon 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