Proceeding of Veterinary and Animal Science Days 2017, 6th- 8th June, Milan, Italy 
 

HAF © 2013 

Vol. IV, No. 1s   ISSN: 2283-3927 

   

 

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Keywords 

Cell culture, Three-dimensional, 

Tissue, Model, Assay 

 

CORRESPONDING AUTHOR 

Stefan Przyborski 

stefan.przyborski@durham.ac.uk 

 

JOURNAL HOME PAGE 

riviste.unimi.it/index.php/haf 

Advanced cell culture technology for 

generation of in vivo-like tissue models 

Stefan Przyborski1,2 

 

 

 

 

1Durham University, Department of Biosciences, Durham, United Kingdom 

2ReproCELL Europe Limited, NETPark Incubator, United Kingdom 

 

Abstract 

Human tissues are mostly composed of different cell types, that are often highly organised in relation 

to each other. Often cells are arranged in distinct layers that enable signalling and cell-to-cell 

interactions. Here we describe the application of scaffold-based technology, that can be used to create 

advanced organotypic 3D models of various tissue types that more closely resemble in vivo-like 

conditions (Knight et al., 2011). The scaffold comprises a highly porous polystyrene material, 

engineered into a 200 micron thick membrane that is presented in various ways including multi-welled 

plates and well inserts, for use with conventional culture plasticware and medium perfusion systems. 

This technology has been applied to generate numerous unique types of co-culture model. For 

example: 1) a full thickness human skin construct comprising dermal fibroblasts and keratinocytes, 

raised to the air-liquid interface to induce cornification of the upper layers (Fig.1) (Hill et al., 2015); 2) a 

neuron-glial co-culture to enable the study of neurite outgrowth interacting with astroglial cells to 

model and investigate the glial scar found in spinal cord injury (Clarke et al., 2016); 3) formation of a 

sub-mucosa consisting of a polarised simple epithelium, layer of ECM proteins simulating the 

basement membrane, and underlying stromal tissues (e.g. intestinal mucosa). These organotypic 

models demonstrate the versatility of scaffold membranes and the creation of advanced in vivo-like 

tissue models. Creating a layered arrangement more closely simulates the true anatomy and 

organisation of cells within many tissue types. The addition of different cell types in a temporal and 

spatial fashion can be used to study inter-cellular relationships and create more physiologically 

relevant in vivo-like cell-based assays. Methods that are relatively straightforward to use and that 

recreate the organised structure of real tissues will become valuable research tools for use in 

discovery, validation studies, and modeling disease. 

http://creativecommons.org/licenses/by-nc-nd/3.0/deed.en


Proceeding of Veterinary and Animal Science Days 2017, 6th- 8th June, Milan, Italy 

HAF © 2013 

Vol. IV, No. 1s   ISSN: 2283-3927 

 

 

 

 
Fig. 1 This figure showcases the potential of 3D cell culture technology and how it can be used to create tissue-like models. The 

histological image reveals the cellular structure of a human skin equivalent. The full thickness of the epidermis is shown resting 

on the underlying dermis. The model is built on the Alvetex® platform that consists of a porous polystyrene scaffold in which 

human dermal fibroblasts are seeded.  These cells produce exogenous collagens to form the dermal compartment. Human 

keratinocytes are then seeded onto the surface of the dermal model that is subsequently raised to air-liquid interface where they 

differentiate, stratify and form a mature epidermis. The layers of cells in the 3D culture model replicate those in the real tissue, 

including the formation of the skin barrier and the surface stratum corneum (Figure courtesy of S.Bradbury, Durham University) 

 

 

References 

Clarke, K.E., Tams, D.M., Henderson, A.P., Roger, M.F., Whiting, A., Przyborski, S., 2016. A robust and reproducible 

human pluripotent stem cell derived model of neurite outgrowth in a three-dimensional culture system and its 

application to study neurite inhibition. Neurochemistry International. 106, 74-84 

Hill, D.S., Robinson, N.D., Caley, M.P., Chen, M., O'Toole, E.A., Armstrong, J.L., Przyborski, S., Lovat, P.E., 2015. A Novel 

Fully Humanized 3D Skin Equivalent to Model Early Melanoma Invasion. Molecular Cancer Therapeutics. 14, 2665-73. 

Knight, E., Murray, B., Carnachan, R., Przyborski, S., 2011. Alvetex®: polystyrene scaffold technology for routine 

three dimensional cell culture. Methods in Molecular Biology. 695, 323-40 

 

http://creativecommons.org/licenses/by-nc-nd/3.0/deed.en