The challenge
A huge hurdle preventing the advancement of preclinical studies is the lack of tough and scalable in vitro culture systems that completely recapitulate what occurs in vivo. Organoids are 3-dimensional (3D) self-replicating miniatures of in vivo tissues and organs and are strong models for ex vivo experimentation in the fields of tissue engineering, disease modeling and drug testing.
The use of an extracellular matrix (ECM), is essential to organoid formation, typically MatrigelTM which stems from a tumorigenic source, and as a result this restricts the translational and scalability of adopting organoids.
The solution
Using synthetic ECM, such as PeptiGel®, for 3D organoid growth will enable scalable and reproducible results. It also opens up the possibility of refining ECM mechanical and chemical functionality in a modular manner to optimize ECM environment for the generation of a broad variety of different organoids.
The science
Single cells or liver crypts were removed from healthy porcine liver tissue and propagated and maintained in liver organoid growth media on five separate PeptiGeIs® Alpha 1-5 (Starter Pack). This enabled the best environment to cultivate gastro-intestinal (Gl) organoids to be determined as an ex vivo model for in vivo systems. The morphology of the subsequent organoids were monitored by utilizing optical microscopy and, as a control, compared to MatrigelTM.
3D organoid growth in a fully synthetic and reproducible ECM like PeptiGels® offers enormous growth potential for pre-clinical studies.
Dr Dammy Olayanju, Senior Research Scientist, Northwick, Park Institute for Medical Research
The results
Hepatic cells were maintained as single cells over the first two days in all PeptiGels®. They then started to cluster and show3D organoid characteristics by day six. By day twelve, all PeptiGels® supported the growth of completely formed organoids within the culture, identical to those grown on MatrigelTM. Intriguingly, PeptiGel® Alpha2 supported the fastest growth of completely formed organoids, which was markedly faster than MatrigelTM (see time-course images taken at x 10 magnification using an inverted optical microscope). Alpha 2 also offered the more stable environment for long term study, lasting at least the one-month in culture investigated here.
Image Credit: Manchester BIOGEL
The future
The PeptiGeI® technology platform offers a reproducible and disease-free ECM for the growth of 3D organoids, which in turn supply more dependable and physiologically pertinent preclinical models for toxicology and pharmacological studies and personally tailored medicine.
About Manchester BIOGEL
Over 15 years ago, Professors Aline Miller and Alberto Saiani at The University of Manchester synthesised a self-assembling oligo-peptide with interesting gelation properties. This work started with a small grant from the University.
Over subsequent years, the team meticulously crafted and studied self-assembling peptides to perfect their platform technology and produce a range of hydrogels ideal for 3D cell culture. In 2014, due to a demand for their materials, our company, Manchester BIOGEL was founded to enable these hydrogels to be readily available to researchers wishing to create new opportunities in the high-growth fields of 3D cell culture, 3D bioprinting and medical devices. Since opening our doors, we have supported scientists in the UK and beyond to create optimal environments for a wide variety of cell types.
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