Board of Ideas

General framework

The current framework is designed to enable responsiveness of all stakeholders to emerging requirements and trends in an agile way. The platform is structured into three levels of operation:

1. A cell culture incubator with thermo and CO2 regulation and IoT functionality for remote access, adjustment and monitoring of the incubation parameters

2. Medium circulation modules, built as trays which can be installed into the incubator or used as stand-alone units. A circulation module contains a pair of syringe pumps (accesed via interface or remotely) and slots for additional components used in the experimental set-up.

3. Customised perfusion components, including tubing, bubble trap, culturing container, etc.

The system can be assembled and customised by modifying existing and/or introducing additional components, such as sensory or stimulatory modules. An outline of possible use-case based development is summarized in the table below:

Level of complexity	Tool development	Possible use-cases	Biological relevance
1	Tray module (in current state) as a standalone unit for controlled perfusion (Wi-Fi, offline control, data logging) used within a standard cell culture incubator. Connectors for use with conventional culturing dishes (petri dish, multi well plate, cell culture flasks).	2D cell cultures (myosatelite cells, striated muscle fibers, fibroblasts, adipocytes, MSCs). 3D cell cultures in scaffolds with large surface area (porous woodpile structures, spheroids, etc.). Stimulation with chemical cues (growth factors, hormones, etc.), which can be added to bulk medium or in intervals and removed through perfusion.	Studying proliferation, differentiation, development and viability of cells under dynamic conditions (mechanical cues, enhanced nutrient gradients, etc.). Cell culture in stable conditions, without cell disturbance (until confluency). Studying the impact of continuous and pulsed chemical stimulation.
2	Tray module with bubble trap and custom culture dish, adapted for a specific scaffold design. Used as a standalone unit within a cell culture incubator.	3D cell culture with a specific scaffold design spatially stabilized, e.g. with internal vasculature, auxetic structures in expanded form, etc. Implementation of external microsensors of ‘basic’ culture parameters (O2, CO2, pH), determining sensor feasibility	Increasing possible scaffold and tissue thickness with high cell viability Prevent or guide scaffold deformation during development Studying the correlation between environmental parameters of the scaffold microenvironment and tissue development
3	Sensory upgrade for tray (based on experimental results of feasibility study) Stimulatory upgrade for tray (electrical stimulation, pulsed perfusion, stretching/compression in elastic culture chamber)	Cultivation of well defined bioscaffolds under complex conditions, that can include electrical (affecting voltage gated channels), mechanical (shear forces due to flow, stretching or compression of the sample) stimuli or both.	Study the impact of scaffold stimulation on tissue development: cell proliferation, differentiation, viability and morphology, etc. Stimulation of muscle fibers during development has been shown to be beneficial in vivo, as well as in vitro.
4	Incubator with CO2 and temperature regulation, central computing and communication unit. Tray modules with individually adjustable flow parameters and temperature fine-tuning. Other modules (sensory, stimulation).	Performing multiple experiments simultaneously with long-distance regulation, monitoring and data logging.	Simultaneous study of several experimental parameters, mentioned above.

Ideas for specific upgrades

Feature	Description	Parameters to consider
Vision system	Compatible microscopy module for monitoring of live cell proliferation and morphology.	illumination magnification working distance depth of field connectivity data collection
Chemical sensors	Monitoring of environmental parameters and culture related changes. Depending on circulation assembly, these can be measured before and after the culturing container. Possible sensors include: pH dissolved oxygen aminoacids growth factors waste products	connectivity data collection probe size compatibility with aseptic workflows (optical sensors)
Stimulation systems	To accelerate cell proliferation and tissue development, sample stimulation can be integrated. Possible mechanisms of stimulation include: mechanical stimulation (pulsed flow, sample deformation, etc.) electrical stimulation chemical stimulation	connectivity mechanism of transduction compatibility with aseptic workflows