Abstract
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.
Highlights
Tissue engineering (TE) for regenerative medicine represents a challenge for the biomedical field with a wide-range of applications, including repair, replacement and regeneration of damaged tissues and organs [1]
The individual cells lost viability after four days of growth and the spontaneously occurring clusteroids grew at much lower rate than the pre-formed clusteroids by our method. These results indicate that HaCaT cells grow well only when successfully attached to a solid substrate or another scaffolding material, as well as by attaching to their neighbouring cells in the clusteroids
In this study we developed a rapid formation method to culture keratinocyte clusteroids by encapsulation of HaCaT cells in a water-in-water Pickering emulsion composed of polyethylene oxide (PEO) as the continuous phase and dextran (DEX) as the dispersed phase
Summary
Tissue engineering (TE) for regenerative medicine represents a challenge for the biomedical field with a wide-range of applications, including repair, replacement and regeneration of damaged tissues and organs [1]. A lack of available organs for transplants or tissue reconstruction slows down recovery of multiple patients. Major risks of these procedures include infection or rejection [2]. TE usually needs three components: Cells, scaffolds and growth factors [3]. Cell culture allows better results as the microenvironment is represented more accurately [4]. Cells usually interact with each other in the extracellular matrix (ECM), a non-cellular scaffold, which plays an important role in multiple processes such as cell proliferation, differentiation or migration [5,6]
Sign-in/Register to view highlights & summary 
Published Version (
Free)
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call