Challenges in Scaling Up 3D Cell Culture for Industrial Use: Navigating Solutions

We discussed the problems faced in scaling up 3D Cell Culture techniques for commercialization. Indentification of hurdles is followed by inspiring solutions! So, let's look at the innovative solutions!

Advanced Bioreactor Designs for Organoids

Invest in bioreactor technologies tailored for organoids, ensuring scalability without compromising complexity. Advances in perfusion systems and dynamic culture environments, as showcased by Driehuis et al., provide a blueprint for large-scale organoid cultivation. Companies like Corning, Molecular Devices and Merck, recognizing the potential of organoid technology, are investing in bioreactor technologies tailored for organoids. Advances in perfusion systems and dynamic culture environments provide a roadmap for large-scale organoid cultivation.

Innovations in Microfluidic System Design

Focus on developing advanced microfluidic systems tailored for industrial-scale applications. Integrating insights from research by Whitesides and Stone on microfluidic control of cell behavior can guide the design of systems that balance precision and scalability. Companies like Dolomite Microfluidics and RainDance Technologies are at the forefront of developing advanced microfluidic systems tailored for industrial-scale applications. Integrating insights from these companies can guide systems design that balance precision and scalability.

Technical solutions to drug adsorption can be overcome by modification of PDMS layers by coating them with inert polymers or investigating treatments such as with gas plasma or UV light, or investigating the mathematical modeling of adsorption to create algorithms that can account for adsorption and inform experimental design.

Bioprinting Approaches

Bioprinting technologies provide several solutions in the domain of 3DCC as the bioprinters and microfluidic devices can be installed in a sterile environment, further, cells are being introduced automatically instead of manually in microfluidic culture chambers. Some bioprinting techniques offer direct immobilization of cells in desirable positions, along with complex printing patterns that can be generated. Additionally, better monitoring of the printed cell ratio can also be achieved, thus biomimetic 3D tissue structures can be printed in vitro.

Animal-free Biomaterials

There needs to be a large-scale production and supply of non-animal origin products with low variability when it comes to formulating or deriving the components. Companies namely HumaBiologics, Manchester Biogel, Growdex, etc supply either human or plant-origin biomaterial and bio-inks for the development of 3DCC systems in vitro. The research in this domain needs to be explored more to sustain a long-term supply of easy-to-manufacture and economical biomimetic products.

Cell Line Engineering

Cell sources have been and will continue to be a challenge for these platforms. It is not straightforward to differentiate all tissues from iPSCs, and primary cells from donors or patients can be hard to come by. To help address the challenges of using heterogeneous iPSCs on the platforms, 3DCC scientists can create cell lines with isogenic backgrounds, some incorporating fluorescent biosensors, to monitor cell differentiation into specific cells. Additionally, the use of genetic editing technology such as CRISPR Cas9 can be used to generate series of differentiated cells of different tissues, whose lineage is known. In future, this technology can also allow for the possibility of silencing or activating disease-associated genes, allowing remarkable potential for research using these platforms on many disease states and treatments. Some 3DCC systems use primary cells and/or populate their platforms with commercially available cell lines, such as those for kidney proximal tubule and liver. Others, such as the female reproductive system organ platforms developed by Woodruff and colleagues and detailed by Burdette et al in this issue, for now, rely on a mixture of animal and human primary tissues due to difficulties in sourcing the tissues.

Automation and Robotics in Spheroid Culture

Implement automation and robotics to streamline the cultivation of spheroids on a larger scale. Automated systems, inspired by the work of Eiraku et al. in optimizing neuroepithelial organoids, enhance precision and reproducibility. Implement automation and robotics to streamline spheroid cultivation on a larger scale. In addition to InSphero, companies like 3D Biomatrix are embracing automation and robotics to streamline spheroid cultivation on a larger scale. These technologies enhance precision and reproducibility in 3D cell culture processes.

Interdisciplinary Collaboration

Foster interdisciplinary collaborations between engineers, biologists, and bioprocess experts to address the challenges of scaling OoC systems. The collaborative efforts of researchers, as seen in the work by Huh et al., demonstrate the potential for translating microphysiological systems to industrial settings. OoC companies, such as Emulate and TissUse, showcase the potential for translating microphysiological systems to industrial settings. Additionally, 3DCC researchers and institutions need to collaborate with industries such as ThermoFisher Scientific, HiMedia, CELLnTEC, STEMCELL Technologies, Corning, etc. to formulate universal media for multi-cell or multi-organ systems.

Scaling up of reliable manufacturing processes for 3DCC systems

The majority of early 3DCC system designs are custom and made on-site at development organizations, where the expense and availability of labor and manufacturing supplies constrain production. Hence, before scale-up can take place, academic labs should concentrate on early quality control of the chips made internally to guarantee reliability and reproducibility. This entails meticulously compiling standard operating procedures for the design and fabrication of chips and?creating clear?quality control procedures that other manufacturers or laboratories can easily follow.

Most academic laboratories are not equipped to scale up 3DCC production, so to mass-produce chips, spin-off or start-up companies or partnerships with manufacturing companies must be formed. It would be very helpful at this point for all manufacturers to conform to Good Manufacturing Practice guidelines, like those published by the FDA. It covers many issues, such as equipment verification, process validation, sanitation and cleanliness of manufacturing facilities, and suitable personnel training. This guidance is not required for OoC manufacturing because it is intended to assure the safety and reliability of manufacturing processes for foods, medications, and medical devices; however, it would still set excellent standards for the dependability of chip production in all industries and contribute to a general increase in system confidence.

Additionally, since this is essential for preclinical toxicology testing and has been linked to high rates of drug development attrition, care should be taken to ensure that all biological assays are created on chips following good laboratory practices. This will help to increase end-user confidence in the fidelity and reliability of mass-produced platforms. Furthermore, independent "qualification" labs, akin to the European Union Reference Laboratory for Alternatives to Animal Testing or the NCATS Tissue Chip Testing Centers, are required to test OoCs and their application with accessible cell types. The term ‘standardization’ brings new challenges concerning what ‘standardization’ means for technical, analytical or biological aspects of 3DCC systems such as OoC. So, ‘performance standards’ should be established for the analytical validation and biological qualification of OoCs. To this end, the deposition of technical, analytical and biological data into a 3DCC systems database will help set some of the standards, reducing the need for users to develop their methods, assays, and analytical methods. At the same time, many government-funded researchers are working with regulatory and industrial end users to evaluate what should be considered accepted metrics that are translatable to other laboratories and applications.

Increasing throughput

Most complex non-organoid tissue chips are currently of very low throughput, where only dozens of replicates (at most) can be performed at any one time. Consequently, during the early stages of drug discovery, at which time many thousands of potential hits can be identified in a short time frame through standard high-throughput screening assays, the use of such chips is likely to be considered cost- and time-prohibitive for pharmaceutical companies at present. Technological advances to create more automated, miniaturized 3DCC systems that can become turnkey technologies for facile use will be crucial to increasing throughput and the number of replicates per platform.

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References

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Author- Deepa Chaturvedi , PhD Scholar, Nanomedicine Research Group, ICTMumbai .