Developing an Open Source Imaging-Driven Multifunctional Bioplotter (IDMB) for next generation in vitro modelling

Lead Research Organisation: King's College London
Department Name: Craniofacial Dev and Stem Cell Biology

Abstract

Cell culture experiments offer unique advantages when trying to study biological mechanisms, which mainly come down to the ability to manipulate both the key cell players and their environment within an experimental set-up, in ways that in vivo experiments would not otherwise allow. This increased control arises from the fact that cell culture systems are inherently less complex than their in vivo counterparts. This reduction of complexity is both an advantage and a crucial limitation of these platforms, and in recent years we have witnessed a plethora of new technologies and tools introduced to increase complexity of cell culture experiments.
Amongst these: stem cell differentiation and reprogramming to obtain the right "key players", live imaging techniques to monitor their dynamic behaviour and interactions, and even bioengineering techniques to control their environment in both 2D and 3D. Thanks to these we can now start to perform more complex experiments that better recapitulate the biological processes we want to study, but we are still limited by our technical capacity to:
i) reliably construct complex arrangements of cell with determined architecture,
ii) directly control and observe the interactions between different cell types and extracellular matrix (ECM), both across space (i.e. their relative position) and time (i.e. how their behavior changes across different time-points during the experiment)
iii) effectively manipulate these complex cultures, which inversely scales with complexity itself
iv) efficiently separate relevant subpopulation of the complex cultures for analysis without losing information on their position and interactions.
To overcome these limitations and study dynamic biological complex systems like neural circuits, developmental niches, tumour microenvironments, or bacterial biofilms we need new platforms that allow us to first construct complex architectures of live cells and ECM, then influence their behaviour & composition in a dynamic way, and finally separate them into relevant subpopulation for analysis. And, as biological systems are dynamic in nature, we need to achieve all of this, while monitoring the behaviour and changes in the composition of the culture.
These capabilities currently do not exist within a single integrated platform, but the technologies necessary are available in the form of i) live imaging platforms, ii) cell-positioning tools, iii) bioplotters or 3D bioprinter and iv) microfluidic-based cell sorters. Particularly, the current technological gap is represented by the fact that while commercial 3D bioprinters and bioplotters can construct complex cultures, they do not allow to acquire live microscopy data, and cannot function as dynamic manipulation tools as they construct cultures without direct feedback. And while live imaging systems that can observe their dynamic behaviour are available, no single platform allows to directly construct or manipulate bioprinted live cell constructs while at the same time acquiring live data on the cell behaviour.
The aim of this project is to capitalise on these different technologies and integrate them within one single prototype platform, that we have named Imaging-Driven Multifunctional Bioplotter (IDMB). This system will be an open-source platform, adaptable to any biology lab and will integrate the functionality of live imaging systems and cell bioplotters/manipulation systems to offer the possibility of plotting live cells and ECM in complex arrangements, as well as manipulate and analyse these complex cultures, in a dynamic way, guided directly by the imaging data. During this project we will construct and optimise a fully functional prototype of the IDMB system and obtain key proof-of-principle data of its application to in vitro modelling.

Technical Summary

This project aims to create a proof-of-principle prototype that integrates the functionality of live imaging systems and 3d-bioplotting within one integrated open-source platform that will allow researchers to construct and manipulate complex in vitro cultures in a dynamic and responsive fashion; performing more complex and relevant in vitro modelling experiments.
This system will be designed to study dynamic complex biological systems: neural circuits, developmental niches, tumour micro-environments, or bacterial biofilms. Allowing, first construction of complex live cells architectures and ECM in complex patterns, then influence their behavior & composition in a dynamic way, and finally separate them into relevant sub-population for analysis. It will achieve all of this, while monitoring the behavior and changes in the composition of the culture using an integrated high-content imaging system.
Currently no single academic or commercially available platform exists that combines all these functionalities within one instrument but the tool and technologies necessary are available and mature. Particularly, the current technological gap is the lack of dynamic manipulation capabilities of 3D bioprinters and bioplotters, which can currently construct complex cultures, but do not allow acquisition of live microscopy data, and cannot therefore be used to monitor and respond to the behavior of the complex cultures being created.
The prototype system, which we have named Imaging-Driven Multifunctional Bioplotter (IDMB) will be an open-source platform adaptable to any biology lab and will offer the possibility of plotting live cells and ECM in complex arrangements, as well as manipulate and analyse these complex cultures, in a dynamic way, guided directly by the imaging data. During this project we will construct and optimise a fully functional prototype of the IDMB system and obtain key proof-of-principle data of its application to in vitro modelling.

Planned Impact

Our main goal is to obtain proof-of-principle for a versatile, open-source bioplotting and cell manipulation system that will allow any biology lab to simultaneously increase complexity and control of in vitro modelling in a way that was before unobtainable without employing multiple large expensive pieces of equipment. With this system, any lab will be able to construct complex in vitro cultures with live cells and matrix, dynamically alter their composition or behaviour and perform correlative live functional imaging together with transcriptomic analysis of defined sub-populations.
We anticipate that this platform will have considerable the impact across different fields of biology and bioengineering (see Academic Beneficiaries), as it represents a novel disruptive tool that integrates multiple functionalities and responds exactly to the arising need in every field of biology to i) obtain more complex in vitro models and ii) manipulate/analyse these complex cultures in a dynamic way.
Moreover, we aim this platform to be completely open source in both hardware and software packages - taking example from successful developments in microscopy such as the Open Microscopy environment and micro-Manager/ImageJ. This will ensure maximum applicability across different labs and allow for this platform to be further modified and optimised for specific experiments within different fields directly by the adopting labs once it is optimised. We have provided within the Case for Support concrete examples of this diverse applications, beside the proposed demonstrations of the platform capabilities with neuroscience experiments, and identified key labs that would be potential early adopters (see Letters of Support)
Moreover, we have also identified industry partners that have an interest in developing or applying the functionality of this platform in collaboration with the investigators (see Pathways to Impact and Letters of Support)

Publications

10 25 50