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Project cofinanced by REACT-EU to mitigate the effects of the pandemic crisis

Project Title: „Enclosed lung on a chip system development suitable for respiratory disease research in BSL3 and microgravity systems. (LoCBox)”

Funding: European Regional Development Fund (ERDF), Measure “Support for applied research”

Project No.:

Period: 1 January 2022 – 30 November 2023

Project costs: 500 000.00 EUR

Principle Investigator: Prof. Arūrs Ābols

Cooperation partner: SIA”Cellboxlab”

Project summary:

This project aims to overcome technical challenges when working with organ on chip models in biological safety level 3 (BSL3) environment and microgravity by developing enclosed test system capable of cultivating cells and be compatible with BLS3 workflow and microgravity systems. Lung on chip model will be tested with pseudoviruses, SARS-Cov-2 and in microgravity device as a proof of principle study.  The following research activities are planned to be carried out during the project:

  • Enclosed system development (LoCBox)
  • LoCBox prototype testing with pseudoviruses
  • LoCBox prototype testing with SARS-Cov-2
  • LoCBox prototype testing with microgravity device
  • Project management and result dissemination

The main outcomes of the project are: two prototypes for lung on chips cultivation and experiments in biological safety level 3 environment and microgravity. We expect that this project will lead to availability of state of art test model and put Latvian Biomedical Research and study center in leading position in technical capabilities for cutting edge research of coronavirus and COVID-19

Information published 03.01.2022.

Progress of the project:

1 January 2022 – 31 March 2022

During this reporting period, we have integrated and optimised pumping solution for the organ on chip cultivation. We have made few iterations of the aluminium holder that holds the pump in place and interfaces with the liquid storage units, currently awaiting the final iteration before placing an order for a small batch. Next, we produced the first batch of COC/OSTE hybrid polymer organs on a chip for lung on a chip development for testing with SARS-COV-2 pseudoviruses. We have improved the OSTE/COC filling process by adjusting the device designs, yield improvement data will be calculated once a larger batch has been tested, but it seems >90% yield within the device is expected. Additionally, during this period, we engaged in public dissemination of project topic in student council of Riga Technical University organised online interview in Spiikiizi studio, titled “What if?”.

Information published 31.03.2022.

Progress of the project:

1 April 2022 – 30 June 2022

During this reporting period, we produced additional COC/OSTE hybrid polymer organs on a chip devices. We have established several lungs on a chip from primary cell cultures with air-liquid interphase. CAD designs, as well as physical prototypes, of BSL3 compatible micropumps have been developed. Cellbox Labs organised a price survey for materials necessary to build a custom BSL3 compatible enclosure. CAD design process of BSL3 compatible enclosure is on-going.

Information published 30.06.2022.

Progress of the project:

1 July 2022 – 30 September 2022

During this reporting period, we developed additional lungs on a chip models from primary cell cultures with air-liquid interphase to quantify ACE2 receptors for pseudovirus infection experiments. Protocol for immunohistochemistry of ACE2 quantification was optimised. Additionally, we have ensued continuous multi-day pumping solution testing to obtain calibration graphs of volume flow rate for the innovative pumping unit. CAD design work for the BSL3-compatible cell culturing box is on-going.

Information published 30.09.2022.

Progress of the project:

1 October 2022 – 31 December 2022

During this reporting period, Cellboxlab have produced additional COC/OSTE hybrid polymer organs on a chip device. Additionally, we had to optimise lungs on a chip models from primary cell cultures with air-liquid interphase to improve immunofluorescence protocols for these models. We continued to work on development of testing units for both BSL3 and microgravity. Cellboxlab also started to prepare invention disclosure form for patent applications.

Finally project supervisors participated in Radio Radio broadcast – The known in the unknown Preclinical studies – organs on a chip and laboratory mice, where they explained to the general public in popular scientific language about technology they are developing.

Information published 30.12.2022.

Progress of the project:

1 January 2023 – 31 March 2023

During this period CellboxLabs produced 20 standard design OOC devices for this project, as well as 20-reduced footprint devices for use in microgravity culturing. Furthermore, Tasks 1.2 and 1.3 have been concluded, thus allowing to start OOC device culturing in BSL3 and microgravity environment.

LBMC started to work on SARS-COV-2 pseudovirus experiments, by establishing models expressing ACE2 receptor. Established workflow and protocols will be used in BSL3 experiments. Additionally, simple experiments of microgravity effect on static cell models have been started that will be used in comparison to Lung on a chip model in microgravity.

Furthermore, CellboxLabs took part in two seminars organized by LBMC and Riga Technical University. The first seminar was on the role of biomedical research in the knowledge economy and biotech start-up success stories, while the second seminar was focused on the transition from university to industry. During these seminars, CellboxLabs presented their project ideas and direction to a wider scientific and industry audience. Finally, LBMC presented results of current progress at international conference.

Information published 31.03.2023.

Progress of the project:

1 April 2023 – 30 June 2023

During this period, we produced additional 10 standard design OOC devices for this project. Furthermore, BSL3 compatible OOC cultivation box has been tested in BSL3 conditions. Pseudovirus infection protocols has been optimised in developed system since 24 hours was not enaught. In parallel Microgravity setup has been tested with microfluidic pumps developed by CellboxLabs and first cell growing experiments have been performed to optimise workflow.

Furthermore, CellboxLabs took part at one of the most popular Latvian Radio stations series: “Mind will save the world” on current and future role of organs on chip. Finally, LBMC and CellboxLabs presented results of current progress of the project at 2nd Microphysiological systems world summit at Berlin.

Information published 30.06.2023.

Progress of the project:

1 July 2023 – 30 September 2023

During this period, we performed Lung on a chip model experiments in microgravity with and without cisplatin in comparison to normal gravity, to evaluate dynamic 3D cancer model cisplatin resistance in microgravity. Additionally, infection experiments with SARS-COV-2 pseudo virus on Lung on a chip and lung on inserts have been carried out for comparison. Furthermore, first live SOSR-COV-2 virus infection experiments on Lung on a chip by implementing BSL3 compatible pumping solutions have been performed.

Finally, CellboxLabs CEO has an interview about technology in company in journal Forbes, CTO visited Rigas Technical University Engineering school, were he explained our technology to pupils and project supervisor Arturs Abols provided an interview to startin.lv to inform industry and journal Medicus Bonus to inform medical doctors about organ on a chip technology and application within this project.

Information published 02.10.2023.

Progress of the project:

1 October 2023 – 30 November 2023

During this pivotal period, we made significant progress in testing the prototypes developed through this project. A major breakthrough was the successful testing of the LocBox prototype, designed to be compatible with Biosafety Level 3 (BSL3) conditions. This prototype was rigorously tested using a SARS-CoV-2 pseudovirus containing the GFP gene. We developed a comprehensive workflow for utilizing the prototype under BSL3 conditions. Building on this success, we proceeded to test the prototype with live SARS-CoV-2 virus, further demonstrating its efficacy and safety.

Another innovative achievement was the testing of a LocBox prototype designed for microgravity conditions. We explored the impact of microgravity on a lung cancer-on-a-chip model using A549 cell lines, specifically focusing on drug resistance, such as to cisplatin, under continuous flow conditions. These tests not only confirmed the effects of microgravity on cancer models, as previously reported in literature, but also revealed significant differences in drug response under microgravity when comparing static to continuous flow conditions.

Our project’s findings and innovations were highlighted at 4 international conferences through both poster and oral presentations. Additionally, two research articles detailing our project’s results have been prepared and submitted for publication. In a significant move towards commercialization, a patent application was submitted by our project collaborator, CellboxLabs, underscoring the practical applications of our research.

Moreover, our commitment to public engagement was evident as we participated in 17 events aimed at disseminating our findings to the broader community. With all project tasks accomplished, as documented in the detailed reports and the final project report, we are proud to confirm the successful completion of this ambitious project.”

Information published 30.11.2023.