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Additive Printing for Cell-Based Analysis

Periodic Reporting for period 3 - ANAPRINT (Additive Printing for Cell-Based Analysis)

Reporting period: 2017-11-01 to 2018-10-31

Context: Mission. ANAPRINT is building better tools for drug discovery, using a combination of additive printing and cryopreservation technology. The tools are advanced human cell models in 3-dimensional (3D) structures, which are used by industry to identify candidate therapeutics and assist their preclinical development. These models also play a critical role in assessing the biological safety of materials ranging from new drugs to environmental pollutants.

Context: Technology. ANAPRINT is realising its technical goals by solving the challenges of applying hard engineering to soft biological materials. Printing pressures and print speeds must recognise the fragility of living cells, and should preserve the cells’ ability to reinstate in culture the tissue-like functions they displayed in vivo. ANAPRINT exploits a decade’s AvantiCell Science experience in cell-handling to stabilise cells prior to printing, deliver them into the printer in a controlled, reproducible manner, and protect them during crucial post-printing equilibration in culture. Alongside this, the 3D scaffold essential for the cells’ organ-like function must be printed in a defined architecture using chemistries which do not compromise incoming or already-resident cells. ANAPRINT must deliver both these outcomes in a repeatable process adapted to the multiwell culture plates used by industry.

Context: Innovation: ANAPRINT will meet market demand for 3D cell models which improve the predictive value of preclinical drug-discovery. Predictive value lies in reproducing, in the cell model, the intracellular processes that make a tissue function and which malfunction in disease. This has become increasingly important as disease intervention targets aberrations in specific, intracellular molecular signals. Without assistance from an in vivo-like printed architecture, tissue-like cell function is not achievable. The paradox is that making such architectures is technically-challenging, laborious and often inconsistent: consequently, industry uptake of 3D cell models has been low, despite their undoubted analytical superiority.

Context: Market. ANAPRINT technologies address a multibillion-euro global market for cell-based analysis. Its technology solutions map onto key market trends towards 3D cell models, use of tissue-reflective cell types, and the delivery of such technology in kit form. Adoption of additive manufacturing as a technical solution reflects wider industry investment in “bio-printing” to produce biological architectures.

Conclusion of the action: Work carried out under the ANAPRINT project has realised the development of a cryopreserved 3D hepatic organoid model for hepatotoxicity testing. Additional technology gains and interim commercial products have included upscaled 3D bioprinting hardware and methodologies, human organoid production and sales, integration of cryopreservation technology and measurement of intracellular molecular readouts in 3D.
"Work performed in the first project period identified additive printing parameters giving optimal quality and throughput. This activity, conducted under two work packages, first established conditions for precision printing of bio-scaffold material and viable cells, and identified constraints on bio-scaffold printing speed and scalability of bio-scaffold additive manufacturing along with their technical solution. During the second and third reporting period, the predictive value and technical performance of printed cell-based assays was defined.

Throughout, protocols have been developed for integration of cell cryopreservation into the manufacturing process: cryopreservation experiments tested utility of conditions suitable for incorporation into a scalable manufacturing process. Further, during the second work package, a series of experiments established working parameters across a hierarchy of cell-product architectures, from printed scaffold and culture medium presentation through to shipped-product packaging/containment. In this way, factors critical to the down-stream maturation of 3D cell models into customer-friendly products were identified, ensuring that further elaborations of the cell models are de-risked in terms of their compatibility with the ultimate concept of a cryopreserved, short lead-time cell-based analysis product.

At the time of project conclusion, ANAPRINT has developed a printed 3D hepatic organoid cell model, in 96 well plate format, for hepatotoxicity testing. This ""plug and play"" assay kit development has been the culmination of the project which has seen many technology gains for ACS, including proprietary cryopreservation technology and bioprinting of cells within biological scaffolds.

Launch of products and services began in Period 2 with a 2D “demonstrator product” showcasing the Cryotix cryopreservation technology. This was followed by the soft launch of human organoid isolates in vial format during the first half of Period 3. The fully-mature ANAPRINT product, using bio-printing to produce plated 3D organoids and Cryotix to freeze them in situ, with promise of 100% recovery into culture, differentiation into mature hepatocytes, and analytical utility is expected to be market ready within the coming months."
Expected Project Outcome. The project objectives remain in place as printing and cryopreservation methodologies are progressively realised. Results achieved at the end of the project are expected to supersede existing products in three ways: 3D-enabled cell performance, consistency of 3D construction (and resultant analytical reproducibility) and convenience of use (“plug and play” utility).

Progress beyond State of the Art. ANAPRINT addresses not only cell model construction but also its user-friendly presentation, by juxtaposing additive manufacture with in situ cryopreservation of cell models at ultra-low temperatures. This combination of leading-edge technologies is expected to bring about a step-change in industry practice, deliverable a cost-effective cell-based analysis solution to an existing, receptive customer base, whilst making advanced analytical technology accessible to new end-users previously unable to adopt and benefit from cell-based analysis as an in-house technical option.

Socio-economic Impact. Better-informed selection/de-selection of preclinical drug candidates enabled by project outputs will produce drug-development savings estimated in $100Millions per successful drug launch, savings which can transform the pharmaceutical industry. Additionally, operational cost savings will provide an immediate incentive to potential customers, by eliminating need to invest in specialised, labour-intensive model construction, with minimal lead time to analysis, and reduced analytical repetition due to inherent model consistency. AvantiCell commercial business is predicted to multiply post-project, with forecast company growth by 30-50 staff for product marketing/manufacturing after 5-7 years.

Societal Benefit. Societal benefit will arise from cost-efficiency gains in the development of new therapeutics, which should deliver major healthcare benefits on a global scale through downward drug price pressure. ANAPRINT outputs are ethically-attractive. 3Rs-compliant drug discovery and less reliance on animal testing are associated with environmental benefit from low infrastructure footprint and low wastage (long shelf-life, short lead-time products).
Human 3D Cell Model Printed in Multiwell Culture Plate
Human 3D hepatic organoid cultures