Scientific posters

Index

Exosomes

PODS® Sustained Release Growth Factors

Cell culture media

 

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Exosome poster brief abstracts 

1. Exo-spin™ mini-HD columns for isolating high-quality exosomes (FSEV 2019) 

Exosome research plays an increasingly important role in therapeutics and diagnostics. Therefore, efficient and reliable methods for isolating exosomes are required. Among other methods, size exclusion chromatography (SEC) is widely used for exosome isolation from all biofluids, as it demonstrates highly pure and concentrated samples. Moreover, SEC can be performed under different protocols, including centrifugation and gravity flow. Under gravity, one sample can be collected into fractions and analysed for a complete high-resolution fractionation profile. In this poster, we highlight the Exo spin™ mini HD kit as a novel tool to isolate exosomes from human serum as an example.

2. A validated technique for exosome isolation from cerebrospinal fluid (ISSCR 2019)

Exosome isolation from cerebrospinal fluid (CSF) is commonly investigated to discover potential protein candidates for diagnostics in central nervous system (CNS) diseases. However, the isolation step is still challenging and an efficient method for isolating exosomes from small volumes of CSF needs to be identified. In this poster, we highlight the Exo-spin™ exosome isolation kit as a novel tool to help exosome isolation in neurodegenerative disease research.

3. Flexible system for isolation of exosomes from small volumes of biofluids (UKEV 2018)

Exosomes can pass the blood-brain barrier due to their nanoscale size. This property makes exosome research attractive for areas such as biomarker discovery for neurodegenerative diseases and also drug delivery. To study these fields, efficient methods for isolating exosomes from small volumes of biofluids such as serum, plasma and CSF need to be identified. Exo-spin™ exosome isolation system was compared to two other precipitation methods using serum, plasma and CSF as starting samples. Exo-spin™ method has been identified as superior. Among other factors, yield, purity as well as structural integrity of the generated samples have been analysed as part of this comparison. Analyses such as NTA, exosome protein to particle ratio, WB and TEM were used to generate comparative data. (TS Martins et al., 2018)

PODS® poster brief abstracts 

1. Armored growth factors transition from 2D and 3D cultures to in-vivo use (ISSCR 2019)

Stem cell culture and differentiation protocols are challenging and generally require a significant amount of time and reagents. One of the main issues is the instability and relatively short half-live of growth factors, which limit their use both in the lab and for transitioning into the clinic. In collaboration with Kyoto Institute of Technology, we developed a novel sustained-release technology which encapsulates growth factors in a protein shell, protecting and preserving their function. PODS (POlyhedrin Delivery System) nanodevices are highly stable and degrade slowly, resulting in a steady release of cargo protein over several weeks. A true platform technology, PODS nanodevices can be applied in different culture systems, in-vitro as well as in-vivo.

2. Sustained release growth factors for stem cell differentiation and organoid culture (LSCN 2019)

Stem cell culture and differentiation protocols are challenging and generally require a significant amount of time and reagents. One of the main issues is instability and the relatively short half-lives of growth factors (GF), which limit their use both in the lab and for transitioning into the clinic. In collaboration with Kyoto Institute of Technology, we developed a novel sustained release growth factor technology which encapsulates growth factors in a protein shell, protecting and preserving their function. PODS (POlyhedrin Delivery System) crystals are produced in insect cells by co-expression of polyhedrin protein and a cargo protein. Highly stable, PODS crystals degrade slowly, resulting in a steady release of cargo protein over several weeks. Here, we outline how PODS crystals can be applied to improve 3D organoid differentiation protocol.

3. Armoured growth factors for tissue engineering (UK Biomaterials 2019)

The inherent instability of recombinant growth factors (GFs), with typical half-lives ranging from minutes to hours, limits their utility in the lab and the clinic. Efficacy may be improved by encapsulation within biomaterials to provide sustained bioavailability over a longer time. However, current delivery systems are limited by protein denaturation and burst release. Refinement of delivery systems to provide sustained release and improved retention may provide therapeutic efficacy at lower doses, improving cost-effectiveness and preventing adverse side effects. PODS (POlyhedrin Delivery System) is a highly durable, crystalline product which encases a GF of interest within polyhedrin protein. The stability of PODS means that crystals degrade slowly, resulting in a steady release of intact, native and functional cargo protein, over several weeks at physiologically-relevant levels.

4. Sustained release of growth factors for therapeutics (UK Biomaterials 2018)

The inherent instability of recombinant growth factors, with typical half-lives ranging from minutes to hours, limits their utility both in the lab and during the transition to the clinic. Furthermore, it is difficult to achieve gradients using standard recombinant growth factors without using special culturing chambers. PODS (POlyhedrin Delivery System) is a highly durable, crystalline product which encases a protein of interest within a polyhedrin protein. This technology exploits the Bombyx mori cypovirus, which encases its mature virion within a polyhedrin protein crystal in order to increase its stability. This stability means that PODS crystals degrade slowly, resulting in a steady release of cargo protein over several weeks. The sustained release mechanism of PODS growth factors can be applied in many ways, from cultivating organoid cultures to functionalising surfaces and scaffolds. We outline how PODS crystals were used to establish a local neurotrophic growth factor gradient. Furthermore, we also demonstrate the therapeutic potential of PODS crystals embedded in a collagen scaffold, using a small animal model of bone repair.

5. A novel approach to protein instability (Milner 2018)

Recombinant growth factors are inherently unstable, with short half-lives limiting their utility in the lab and clinic. PODS (POlyhedrin Delivery System) are produced in cultured insect cells by co-expression of polyhedrin protein with a cargo protein. This encases and protects the protein of interest within a polyhedrin crystal. PODS crystals contain intact, native, and functional protein, are highly durable and extremely stable in storage, and degrade slowly over several weeks to steadily release active cargo protein. The sustained release mechanism of PODS growth factors can be used in many ways, e.g., to cultivate organoid cultures, create growth factor gradients or to functionalise surfaces and scaffolds.

Cell culture media poster brief abstracts 

1. A fully defined system for confluent human pluripotent cell culture in the absence of bFGF

The Pluripro® system, a fully defined medium partnered with fully defined matrix, enables enzymatic, single cell passaging of human pluripotent stem cells (hPSCs). The Pluripro system provides consistency of hPSC phenotype both within and between passages. Following extended passage in Pluripro, hPSCs retain an undifferentiated phenotype, as evidenced by expression of markers for the undifferentiated state, and capacity for in-vitro differentiation into the three germ lineages.