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Fabrication of Angiogenic Sprouting Coculture of Cell Clusteroids Using an Aqueous Two-Phase Pickering Emulsion System

Wang, Anheng; Madden, Leigh A.; Paunov, Vesselin N.

Authors

Anheng Wang

Vesselin N. Paunov



Abstract

Tumor cell spheroids and 3D cell culture have generated a lot of interest in the past decade due to their relative ease of production and biomedical research applications. To date, the frontier in tumor 3D models has been pushed to the level of personalized cancer treatment and customized tissue engineering applications. However, without vascularization, the central parts of these artificial constructs cannot survive without an adequate oxygen and nutrient supply. The formation of a necrotic core into in vitro 3D cell models still serves as the major obstacle in their wider practical application. Here, we propose a rapid formation protocol based on using a water-in-water (w/w) Pickering emulsion template to generate phenotypically endothelial/hepatic (ECV304/Hep-G2) coculture cell clusteroids with angiogenic capability. The w/w Pickering emulsion template was based on a dextran/poly(ethylene oxide) aqueous two-phase system stabilized by whey protein particles. The initial cell proportion in the coculture clusteroids can easily be manipulated for optimal performance. The cocultured pattern of the endothelial/hepatic cells could significantly promote the production of angiogenesis-related proteins. Our study confirmed that cocultured clusteroids can stimulate cell sprouting without the addition of vascular endothelial growth factor (VEGF) or other angiogenesis inducers at a 1:2 ratio of Hep-G2/ECV304. Angiogenesis gene production in the coculture clusteroids was enhanced with VEGF, urea, and insulin-like growth factor-binding protein along with angiogenesis-related marker CD34 levels, also indicating angiogenesis progress. Our aqueous two-phase Pickering emulsion templates provided a convenient approach to vascularize a target cell type in 3D cell coculture without additional stimulating factors, which could potentially apply to either cell lines or biopsy tissues, expanding the clusteroids downstream applications.

Citation

Wang, A., Madden, L. A., & Paunov, V. N. (2022). Fabrication of Angiogenic Sprouting Coculture of Cell Clusteroids Using an Aqueous Two-Phase Pickering Emulsion System. ACS Applied Bio Materials, https://doi.org/10.1021/acsabm.2c00168

Journal Article Type Article
Acceptance Date Mar 9, 2022
Online Publication Date Mar 22, 2022
Publication Date 2022
Deposit Date Mar 22, 2022
Publicly Available Date Mar 23, 2023
Journal ACS Applied Bio Materials
Print ISSN 2576-6422
Electronic ISSN 2576-6422
Publisher American Chemical Society
Peer Reviewed Peer Reviewed
DOI https://doi.org/10.1021/acsabm.2c00168
Keywords Coculture; Angiogenesis; Aqueous two-phase system; Water-in-water Pickering emulsion; 3D cell culture
Public URL https://hull-repository.worktribe.com/output/3953382

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Copyright Statement
© American Chemical Society




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