Prof Stefan Przyborski speaking at BBS 2016 Biennial Meeting

ReproCELL Europe Ltd’s Chief Scientific Officer Prof Stefan Przyborski is giving a public lecture on “Scaffold Cell Culture Technology for Generation of In Vivo-like Tissue Models” at the British Biophysical Society 2016 Biennial Meeting 6th-8th July, University of Liverpool, Liverpool, UK.



Scaffold Cell Culture Technology for Generation of In Vivo-like Tissue Models

Stefan Przyborskia,b
aSchool of Biological and Biomedical Science, Durham University, South Road, Durham DH1 2PJ, UK
bReinnervate ReproCELL, NETPark Incubator, Thomas Wright Way, Sedgefield TS21 3FD, UK

The benefits of three dimensional (3D) cell culture are widely appreciated. More cell-based technologies are now becoming available that enable researchers to preserve the native 3D structure of cells in vitro. These can be broadly divided into three areas: aggregate-based methods; hydrogels and extra-cellular matrices; and inert scaffold-based technologies. Each has strengths and weaknesses and there is no one technology that satisfies all applications. Tissues in the body 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. Recreation of these types of architecture will significantly evolve 3D cell culture to a new level where real tissue-like structures can be generated in vitro for research and discovery. Here we describe the application of Alvetex®, a 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.

Alvetex is a highly porous polystyrene scaffold that is engineered into a 200 micron thick membrane. The scaffold is presented in various ways including multi-welled plates and well inserts, for use with conventional culture plasticware and a novel medium perfusion system. Collectively, this technology has been applied to generate numerous unique types of co-culture model, for example: 1) a full thickness human skin construct comprising primary dermal fibroblasts and keratinocytes, raised to the air-liquid interface to induce cornification of the upper layers; 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; 3) a cancer invasion model where a 3D layer of stromal cells co-cultured by invading colorectal cancer cells; 4) formation of a sub-mucosa consisting of a polarised simple epithelium, layer of ECM proteins simulating the basement membrane, and underlying stromal tissues. These organotypic models demonstrate the versatility of Alvetex technology and the creation of advanced in vivo-like tissue models. Creating a layered arrangement more closely simulates the true organisation of cells within many tissue types. Using such scaffold-based technology also provides the opportunity to control the addition of different cell types in a temporal and spatial fashion, thus building up a model over time, to study inter-cellular relationships.

The ability to maintain and study tissue slices ex vivo is also an attractive application of Alvetex. The highly porous material enhances tissue viability and also acts to hold the sample stable during longterm live cell imaging experiments. This has been successfully applied to neurite outgrowth studies in spinal cord slices and the longterm maintenance of human stem cell derived tissues. It also has major implications for maintaining tissues ex vivo for approaches in personalised medicine.

More sophisticated models are developing as 3D cell culture technology becomes established and accepted as a means of creating 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.