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Fabrication of Human Keratinocyte Cell Clusters for Skin Graft Applications by Templating Water-in-Water Pickering Emulsions

Celik, Sevde; Dominici, Sebastien; Filby, Benjamin; Das, Anupam; Madden, Leigh A.; Paunov, Vesselin N.


Sevde Celik

Sebastien Dominici

Benjamin Filby

Anupam Das

Vesselin N. Paunov


Most current methods for the preparation of tissue spheroids require complex materials, involve tedious physical steps and are generally not scalable. We report a novel alternative, which is both inexpensive and up-scalable, to produce large quantities of viable human keratinocyte cell clusters (clusteroids). The method is based on a two-phase aqueous system of incompatible polymers forming a stable water-in-water (w/w) emulsion, which enabled us to rapidly fabricate cell clusteroids from HaCaT cells. We used w/w Pickering emulsion from aqueous solutions of the polymers dextran (DEX) and polyethylene oxide (PEO) and a particle stabilizer based on whey
protein (WP). The HaCaT cells clearly preferred to distribute into the DEX-rich phase and this property was utilized to encapsulate them in the water-in-water (DEX-in-PEO) emulsion drops then osmotically shrank to compress them into clusters. Prepared formulations of HaCaT keratinocyte clusteroids in alginate hydrogel were grown where the cells percolated to mimic 3D tissue. The HaCaT cell clusteroids grew faster in the alginate film compared to the individual cells formulated in the same matrix. This methodology could potentially be utilised in biomedical applications.


Das, A. A., Dominici, S. R., Filby, B. W., Celik, S. B., Celik, S., Dominici, S., …Paunov, V. N. (2019). Fabrication of Human Keratinocyte Cell Clusters for Skin Graft Applications by Templating Water-in-Water Pickering Emulsions. Biomimetics, 4(3),

Journal Article Type Article
Acceptance Date Jul 8, 2019
Online Publication Date Jul 11, 2019
Publication Date Sep 1, 2019
Deposit Date Jul 15, 2019
Publicly Available Date Jul 16, 2019
Journal Biomimetics
Electronic ISSN 2313-7673
Publisher MDPI
Peer Reviewed Peer Reviewed
Volume 4
Issue 3
Article Number 50
Keywords Tissue engineering; Spheroids; Keratinocyte; HaCaT; Water-in-water emulsions; Pickering emulsions; PEO; DEX; Hydrogels; Alginate
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Copyright Statement
© 2019 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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