M.Sc.

Nießing, Bastian

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  • Publication
    Automated Production at Scale of Induced Pluripotent Stem Cell-Derived Mesenchymal Stromal Cells, Chondrocytes and Extracellular Vehicles: Towards Real-Time Release
    ( 2023-10-10)
    Herbst, Laura
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    Groten, Ferdinand
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    Murphy, Mary
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    Shaw, Georgina
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    Nießing, Bastian
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    Schmitt, Robert H.
    Induced pluripotent stem cell (iPSC)-derived mesenchymal stem cells (iMSCs) are amenable for use in a clinical setting for treatment of osteoarthritis (OA), which remains one of the major illnesses worldwide. Aside from iPSC-derived iMSCs, chondrocytes (iCHO) and extracellular vesicles (EV) are also promising candidates for treatment of OA. Manufacturing and quality control of iPSC-derived therapies is mainly manual and thus highly time consuming and susceptible to human error. A major challenge in translating iPSC-based treatments more widely is the lack of sufficiently scaled production technologies from seeding to fill-and-finish. Formerly, the Autostem platform was developed for the expansion of tissue-derived MSCs at scale in stirred tank bioreactors and subsequent fill-and-finish. Additionally, the StemCellDiscovery platform was developed to handle plate-based cultivation of adherent cells including their microscopic analysis. By combining the existing automation technology of both platforms, all required procedures can be integrated in the AutoCRAT system, designed to handle iPSC expansion, differentiation to iMSCs and iCHOs, pilot scale expansion, and formulation of iMSCs as well as extracellular vesicles and their purification. Furthermore, the platform is equipped with several in-line and at-line assays to determine product quality, purity, and safety. This paper highlights the need for adaptable and modular automation concepts. It also stresses the importance of ensuring safety of generated therapies by incorporating automated release testing and cleaning solutions in automated systems. The adapted platform concepts presented here will help translate these technologies for clinical production at the necessary scale.
  • Publication
    Towards automated CAR-T Cell Manufacturing. Keeping up with Technological Advancement
    ( 2023-05-04)
    Herbst, Laura
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    Erkens, Frederik
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    Hort, Simon orcid-logo
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    Bäckel, Niklas
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    Nießing, Bastian
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    Popp, Georg
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    Franz, Paul
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    König, Niels orcid-logo
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    Hudecek, Michael
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    Rafiq, Qasim
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    Goldrick, Stephen
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    Papantoniou, Ioannis
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    Schmitt, Robert H.
    The AIDPATH project has received funding from the European Union's Horizon 2020 research and innovation programme under grant agreement no 101016909. The material presented and views expressed here are the responsibility of the author(s) only. The EU Commission takes no responsibility for any use made of the information set out.
  • Publication
    Adaptive phase contrast microscopy to compensate for the meniscus effect
    ( 2023-04-08)
    Nienhaus, Florian orcid-logo
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    Piotrowski, Tobias
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    Nießing, Bastian
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    König, Niels orcid-logo
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    Schmitt, Robert H.
    Phase contrast is one of the most important microscopic methods for making visible transparent, unstained cells. Cell cultures are often cultivated in microtiter plates, consisting of several cylindrical wells. The surface tension of the culture medium forms a liquid lens within the well, causing phase contrast conditions to fail in the more curved edge areas, preventing cell observation. Adaptive phase contrast microscopy is a method to strongly increase the observable area by optically compensating for the meniscus effect. The microscope’s condenser annulus is replaced by a transmissive LCD to allow dynamic changes. A deformable, liquid-filled prism is placed in the illumination path. The prism’s surface angle is adaptively inclined to refract transmitted light so that the tangential angle of the liquid lens can be compensated. Besides the observation of the phase contrast image, a beam splitter allows to simultaneously view condenser annulus and phase ring displacement. Algorithms analyze the displacement to dynamically adjust the LCD and prism to guarantee phase contrast conditions. Experiments show a significant increase in observable area, especially for small well sizes. For 96-well-plates, more than twelve times the area can be examined under phase contrast conditions instead of standard phase contrast microscopy.
  • Publication
    High-Speed-Microscopy for Scalable Quality Control in Automated Production of Stem Cell Spheroids for Tissue Engineering
    ( 2023)
    Krieger, Judith
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    Nießing, Bastian
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    König, Niels orcid-logo
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    Schmitt, Robert
    The EU Horizon 2020 project »JointPromise« implies the conception and implementation of an end-to-end automated production platform for three-dimensional joint implants, paving the way for tissue-engineered implants able to regenerate deep osteochondral defects. Spheroid-based implants provide a novel approach in tissue engineering by aggregating progenitor cells into potent microtissues. After the differentiation of cartilaginous microtissues, functional joint implants are assembled via 3D bioprinting to match the complex structural organization of native cartilage tissue. As the automation approach of the project aims to overcome bottlenecks in manual production such as product variability, lack of scalability and high personnel costs, a high-throughput quality control system is crucial for the production of reliable Advanced Therapy Medicinal Products (ATMPs). By establishing not only a technical solution for the full digitization of the cell culture plates but also an intelligent image processing algorithm for the detection of the cell spheroids, relevant process parameters like size distribution and growth curves can be detected. Critical thresholds in spheroid growth are evaluated to minimize risks of carcinogenic tissue formation in vivo as well as to define harvest criteria to prevent inhomogeneous bioprinting results. In order to calculate the required throughput and elaborate optimization potentials of the automated spheroid production, voids in the cultivation vessel or disrupted aggregates due to media changes or transportation are detected. Ultimately, the high-speed-microscopy complies with the requirements of a high-throughput automated cell production platform to meet the rising demand for alternative therapeutic approaches in regenerative medicine.
  • Publication
    Toward Rapid, Widely Available Autologous CAR-T Cell Therapy - Artificial Intelligence and Automation Enabling the Smart Manufacturing Hospital
    ( 2022-06-06)
    Hort, Simon orcid-logo
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    Herbst, Laura
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    Bäckel, Niklas
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    Erkens, Frederik
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    Nießing, Bastian
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    Frye, Maik
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    König, Niels orcid-logo
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    Papantoniou, Ioannis
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    Hudecek, Michael
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    Jacobs, John J. L.
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    Schmitt, Robert
    CAR-T cell therapy is a promising treatment for acute leukemia and lymphoma. CAR-T cell therapies take a pioneering role in autologous gene therapy with three EMA-approved products. However, the chance of clinical success remains relatively low as the applicability of CAR-T cell therapy suffers from long, labor-intensive manufacturing and a lack of comprehensive insight into the bioprocess. This leads to high manufacturing costs and limited clinical success, preventing the widespread use of CAR-T cell therapies. New manufacturing approaches are needed to lower costs to improve manufacturing capacity and shorten provision times. Semi-automated devices such as the Miltenyi Prodigy® were developed to reduce hands-on production time. However, these devices are not equipped with the process analytical technology necessary to fully characterize and control the process. An automated AI-driven CAR-T cell manufacturing platform in smart manufacturing hospitals (SMH) is being developed to address these challenges. Automation will increase the cost-effectiveness and robustness of manufacturing. Using Artificial Intelligence (AI) to interpret the data collected on the platform will provide valuable process insights and drive decisions for process optimization. The smart integration of automated CAR-T cell manufacturing platforms into hospitals enables the independent manufacture of autologous CAR-T cell products. In this perspective, we will be discussing current challenges and opportunities of the patient-specific but highly automated, AI-enabled CAR-T cell manufacturing. A first automation concept will be shown, including a system architecture based on current Industry 4.0 approaches for AI integration.
  • Publication
    Implementation of an Automated Manufacturing Platform for Engineering of Functional Osteochondral Implants
    ( 2022)
    Krieger, Judith
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    Nießing, Bastian
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    König, Niels orcid-logo
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    Mota, Carlos
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    Pointe, Vanessa la
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    Rijt, Sabine van
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    Kondro, Douglas
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    Hiatt, Michael
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    Viellerobe, Bertrand
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    Brisson, Bruno
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    Marechal, Marina
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    Geris, Liesbet
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    Luyten, Frank P.
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    Papantoniou, Ioannis
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    Schmitt, Robert
    The EU Horizon 2020 project »JointPromise« proposes the development and implementation of an end-to-end automated production platform for three-dimensional joint implants, paving the way for tissue-engineered implants able to regenerate deep osteochondral defects. Currently, the manufacturing pipeline consists in manual production processes for microtissue cultivation, harvest and bioassembly into larger implants. In the conceptualizing stage of this project, the manual processes were translated into standard operating protocols (SOPs) and process design criteria like material flow and throughput as well as technical specifications of laboratory devices for an automated performance were elaborated. Spheroid-based implants provide a novel approach in tissue engineering by aggregating progenitor cells into potent microtissues. After the differentiation of cartilaginous microtissues, functional joint implants are assembled via 3D bioprinting to match the complex structural organization of native cartilage tissue. The »JointPromise« platform includes suitable devices for cell and microtissue cultivation, harvest and implant production as well as quality control in an overall layout consisting of according pipetting units, incubator, centrifuge, bioprinter and high-speed microscope. After initiating the platform build-up, a control software for process controlling and monitoring during cell seeding, cultivation and harvest is implemented. Clinical feasibility and efficacy of osteochondral defect regeneration by the produced joint implants will subsequently be proven in large animal models.
  • Publication
    Automated Manufacturing of Microtissue Based Osteochondral Implants: The »JOINTPROMISE« Platform
    ( 2022)
    Krieger, Judith
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    Nießing, Bastian
    ;
    König, Niels orcid-logo
    ;
    Luyten, Frank P.
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    Papantoniou, Ioannis
    ;
    Schmitt, Robert H.
    Over 300 million cases of osteoarthritis were reported in 2017, stating one of the most prevalent chronic joint diseases worldwide characterized predominantly by long-term progressive cartilage and subchondral bone degeneration. Conventional therapy approaches utilize pharmacotherapy mostly for pain relief and at end stage disease treated by whole joint replacement surgery to retrieve some mobility and function. Novel Regenerative Medicine (RM) strategies employing Tissue Engineered implants could enable cure, more than care, of such life-constraining disabilities to meet the rising demand for medical interventions due to an ageing world population. Engineering joint tissue implants for the regeneration of the cartilage-bone unit of the joints remains a challenge due to the complex structural organization and functionality of native joint tissue. The use of microtissue/spheroid platforms has enabled differentiation and maturation of cartilage intermediates and gives hope for the engineering of efficient large-scale implants for osteochondral regeneration. However, there is still lack of enabling technologies for scaling of these approaches and robust manufacturing with end-to-end automation of such advanced therapeutic medicinal products (ATMPs). To allow sufficient scaling, overcome risks of contamination as well as inconsistent product quality in manual production procedures, the automated, GMP-compliant manufacturing platform »JointPromise« is developed. By establishing a robust, large-scale manufacturing process, a reliable microtissue-based product for the treatment of deep osteochondral defects can be generated with suitable productivity. The platform concept is based on the translation of Standard Operating Procedures (SOPs) for microtissue production, harvest and condensation into a sequence of automated process steps. Derived process design criteria and technical specifications result in device requirements for an automated production process. After initiating the conceptualizing stage of the platform design by creating a 2D layout according to the material flow of the translated SOPs, the final arrangement of devices was optimized in the overall 3D CAD model. The resulting production platform model combines all required devices for cell cultivation, microtissue harvest and ATMP production in an overall layout consisting of pipetting units, an incubator, centrifuge, bioprinter and housing for a defined hygienic environment. Following the SOPs, about 28,000 microtissue spheroids can be produced within 21 days of culture out of 1 mL cell suspension per tissue culture plate. To reach the required productivity of around 100 tissue culture plates per implant, the production platform will need to process around 70 L of liquids during seeding and harvest processes and 5 L per cell media change to produce around 2.8M microtissue spheroids in 21 days. The build-up of the »JointPromise« platform is followed by the implementation of the control software COPE (Control Operate Plan Execute, Fraunhofer IPT, Aachen, Germany) for process controlling and monitoring during cell seeding, cultivation and harvest.