Research Project

Morphometry

FAD/NADH real-time imaging

The process of establishing new organoid lines currently suffers from a significant bottleneck during early culture due to a “catch-22” situation; optimisation of culture conditions is needed to grow organoids well, but this requires destruction of many of those organoids in early culture during the optimisation process. The reason for this is that almost all the tests currently in use to measure the viability of organoids require the cells to be broken apart and destroyed.

This project seeks to transfer and embed essential new knowledge into Cellesce to enable the Company to develop novel cell ‘happiness’ assays to allow real-time optimisation of the growth of patient samples as 3D organoids.

Live-cell imaging enables the non-destructive observation of organoid metabolic activity, viability and health from smaller numbers of organoids. This Knowledge Transfer Partnership (KTP) aims to develop a workflow that identifies the optimal growth conditions required for the cost effective, rapid expansion of small amounts of starting material for each new organoid line prior to expansion in Cellesce’s bioreactors.

This project will embed new knowledge into Cellesce in the following three areas:

  1. Non-invasive imaging – Visualisation of the organoid and assessment of its metabolism/health. This is critical when maintaining live organoids in a strictly controlled environment. Any molecule added into the system (such as a fluorescent marker) has the potential to dramatically affect growth and differentiation.
  2. Live cell analysis – Replicating the carefully controlled environment of a tissue culture incubator on a piece of equipment such as a microscope requires expertise to fine tune the set-up.
  3. Image analysis – 3D live-cell analysis of multiple components requires both spatial and temporal correlation of the data produced.

This project will identify optimal microscopy hardware and image analysis solutions required to enable experimental workflows to be implemented in house. The Research Associate will make recommendations to the Board to fulfil this requirement.Knowledge at the technical level will be transferred through interaction of the Associate with colleagues at Cellesce during regular joint meetings. The formal transfer of expertise will be codified in Standard Operating Procedures (SOPs) that will be integrated into Cellesce workflows.
This project started in July 2019 and is due to run until June 2021. It is a collaborative project with Cardiff University supervised by and Dr Pete Watson and Professor Karl Swann within the School of Biosciences.

Funded by: Innovate UK (KTP)