Research Project

Organoid Expansion

Establishing a translational advanced pilot manufacturing ecosystem for future biomedical products

This project is focused on three main tasks. Cellesce provides the knowledge in the bioprocessing technology for the growing and expansion of the organoids. INL provide the micro-fluidics and lab-on- a-chip expertise.

Design and fabrication of a microfluidic device for optimised culture of 3D tumour organoids.

To design a microfluidic device suitable for the culturing and maintenance of organoids on-chip.The optimum size/growth extent of organoids will be first evaluated in vitro. For that, cryopreserved tumour organoids provided by Cellesce will be thawed at INL and seeded in Matrigel according to Cellesce established protocol. After a short recovery, tumour organoids will be microscopically evaluated in order to assess their volume and size. Such parameters will be taken into consideration to accurately design a microfluidic device for optimised expansion of organoids on-chip. Afterwards the microfluidic device will be fabricated and characterised at INL, taking advantage of the available state-of-the-art facilities for nano-fabrication and nano-characterisation.

Evaluation of microfluidic chip performance for culturing tumour organoids.

When appropriate, tumour organoids are transferred to the microfluidic device and embedded in micro- patterned 3D matrices immediately contiguous to a microchannel. The growth and viability of organoids-on-chip will be monitored by microscopy imaging for three to five days. In parallel, input and output flow conditions will be optimised and temperature, pH, nutrient and oxygen supply and waste removal will be evaluated in order to ensure full development of the organoids. When the best culturing conditions have been defined, generation of tumour organoids from original cells inside the microfluidic chip will be pursued.

Development of micro-fluidic co-culture systems for tumour immunology studies.

Building on the findings from the previous tasks, and in the absence of time constraints, increasing the complexity of the microfluidic platform will be attempted to accommodate the presence of more than tissue type. Specifically, this will look at the interactions between tumour organoids with immune cells (T cells and or NK cells) to assess specific killing of cells in vitro. Tumour organoids will be co-cultured on chip with peptide-specific T cells and organoids pulsed with TCR-specific peptide prior to co-culture. Antigen specific killing by T cells and possible T cell infiltration into the epithelium of the organoids will be analysed by microscopy imaging. If possible, experiments with autologous immune cells from cancer patients will also be conducted.