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Vascularized Co‐Culture Clusteroids of Primary Endothelial and Hep‐G2 Cells Based on Aqueous Two‐Phase Pickering Emulsions

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


Anheng Wang

Vesselin N. Paunov


Three‐dimensional cell culture has been extensively involved in biomedical applications due to its high availability and relatively mature biochemical properties. However, single 3D cell culture models based on hydrogel or various scaffolds do not meet the more in‐depth requirements of in vitro models. The necrotic core formation inhibits the utilization of the 3D cell culture ex vivo as oxygen permeation is impaired in the absence of blood vessels. We report a simple method to facilitate the formation of angiogenic HUVEC (human umbilical vein endothelial cells) and Hep‐G2 (hepatocyte carcinoma model) co‐culture 3D clusteroids in a water‐in‐water (w/w) Pickering emulsions template which can overcome this limitation. This method enabled us to manipulate the cells proportion in order to achieve the optimal condition for stimulating the production of various angiogenic protein markers in the co‐cultured clusteroids. The HUVEC cells respond to the presence of Hep‐G2 cells and their byproducts by forming endothelial cell sprouts in Matrigel without the exogenous addition of vascular endothelial growth factor (VEGF) or other angiogenesis inducers. This culture method can be easily replicated to produce other types of cell co‐culture spheroids. The w/w Pickering emulsion template can facilitate the fabrication of 3D co‐culture models to a great extent and be further utilized in drug testing and tissue engineering applications.


Wang, A., Madden, L. A., & Paunov, V. N. (2022). Vascularized Co‐Culture Clusteroids of Primary Endothelial and Hep‐G2 Cells Based on Aqueous Two‐Phase Pickering Emulsions. Bioengineering, 9(3), Article 126.

Journal Article Type Article
Acceptance Date Mar 9, 2022
Online Publication Date Mar 21, 2022
Publication Date 2022-03
Deposit Date Mar 22, 2022
Publicly Available Date Oct 27, 2022
Journal Bioengineering
Print ISSN 2306-5354
Electronic ISSN 2306-5354
Publisher MDPI
Peer Reviewed Peer Reviewed
Volume 9
Issue 3
Article Number 126
Keywords Co-culture; Angiogenesis; Water-in-water Pickering emulsions; 3D cell culture
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Copyright Statement
© 2022 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (

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