Leading voices in the field such as Prof. Dr. Gerhard Gstraunthaler, emer., University of Innsbruck, Austria, Dr. Lothar Steeb, CEO/CSO PELOBiotech, discussed ethical considerations regarding FBS and alternatives like human Platelet Lysate (hPL), serum-free, animal component-free media, xeno-free options, and defined media and the latest developments in translational medicine. Just click here to read more.
Clinical Potential of MSC-EVs and Translational Challenges
Prof. Dr. rer. nat. Bernd Giebel
Human mesenchymal stromal cells (MSCs) are a therapeutically relevant, heterogeneous cell entity with immunomodulatory and pro-regenerative potentials. MSCs mediate a huge proportion of their therapeutic effects via extracellular vesicles (EVs). Connected to several advantages in using cell-free products for the therapeutic setting, MSC-EVs emerged as promising novel therapeutic agents for various diseases, including graft-versus-host disease (GvHD), ischemic stroke, COVID-19 and sepsis.
It is our current mission to optimize the MSC-EV production strategy in a scaled, GMP-compliant manner, and to set up an appropriate quality control platform to translate MSC-EVs into the clinics. One of the challenging aspects in this context is inherited from the MSC field, i.e. contradictory reports on the efficacy of MSC therapies. Not all MSC products mediate therapeutic effects when applied to patients. Similarly, we observe functional differences among independent MSC-EV preparations; even when the same MSC stocks were used as starting material. Thus, to avoid drawbacks as they occurred in the MSC field by failing to show efficacy in a phase III clinical trial for GvHD treatment, it is one of our most important missions to address and appropriately handle the heterogeneity aspect. To this end, we have set up a lentiviral, hTERT-based immortalization strategy and raised MSC lines at the clonal level. EVs released by these clonally expanded immortalized MSCs (ciMSCs) reveal immunomodulatory activities and confer therapeutic activities in vivo. According to our understanding, we thus have fulfilled an essential milestone towards scaled and standardized production of MSC-EV-based therapeutics.
The Long and Winding Road to Serum Alternatives
Prof. Gerhard Gstraunthaler, Ph.D., emer
The supplementation of cell culture media with fetal bovine serum (FBS) is routine practice, however, FBS bears a number of disadvantages:
- an animal-derived component of unknown composition,
- seasonal and geographical lot-to-lot variability,
- safety concerns in terms of endotoxins, mycoplasma, viral contaminants or prion proteins, and
- unpredictable shortages in global supply. Most serious are
- ethical concerns on animal welfare regarding blood collection from unborn bovine fetuses.
In light of the above mentioned scientific, consistency and reproducibility, ethical, and supply challenges, several strategies have been developed to reduce or replace FBS in cell culture media and the search for alternatives to FBS has become a major goal in the field of cell and tissue culture research.
Most recently, releasates of activated human donor thrombocytes (human platelet lysates, hPL) are one of the most promising alternatives to FBS. The rationale behind the use of hPL is the clotting process itself, which includes the activation of thrombocytes and the exocytic release of α-granule factors, respectively. Platelet α-granules contain an array of growth factors, required for wound healing and tissue regeneration, that have also been identified as being essential for cell attachment, growth, and proliferation in vitro.
The source of origin for hPL is expired human donor thrombocytes, which is a well-recognized, ethically approved, safe, and clinically tested, high-quality product. With hPL, bulk human thrombocytic growth factors are added to basal culture media, providing a human-based, xeno-free culture system.
Cardiac organoids: Scaling up for preclinical studies
Prof. Dr. Kurt Pfannkuche
Cardiac organoids (COs) derived from human induced pluripotent stem (iPS) cells contain the major cell types of the heart. COs are therefore promising for transplantation in experiments to restore infarcted heart tissue. Today, iPS cell technologies are moving from basic research to (pre-)clinical applications. Therefore, protocols need to be adapted to be compatible with GMP manufacturing to streamline the clinical translation of research results. In this presentation, a technology to generate COs in stirred bioreactors under defined FCS-free conditions will be presented and new developments in bioreactor technologies will be discussed.
Human adipocyte spheroids: Tools to study obesity-related questions in a more in vivo-like environment
Dr. rer. nat. Anita Wagner
In vitro model of human subcutaneous adipocyte spheroids to investigate mitochondrial dysfunction and mitochondria activating compounds
Mitochondrial abnormalities drive obesity-related adipose tissue dysfunction and metabolic complications. Understanding adipocyte mitochondrial dysfunction and identifying activating compounds may aid in treating obesity-related disorders. A major hurdle is the lack of a physiological in vitro model for studying mitochondria in human adipocytes. Our goal was to establish a human white subcutaneous adipocyte spheroid model to characterize mitochondrial metabolism in obesity-relevant conditions and drug exposure. We utilized preadipocytes from two donors with different BMIs, cultured as undifferentiated or differentiated spheroids. Differentiated spheroids show increased lipid accumulation, gene expression, mitochondrial respiration, and adiponectin secretion. They responded well to insulin and β-adrenergic stimulation. Lipid treatment reduced mitochondrial function, while other components improved it without affecting lipid accumulation. Our model replicates in vivo adipocyte function and enables precise measurement of mitochondrial function under various conditions, facilitating drug screening and advancing obesity-related research.
Humanized Multi-Organoid Disease Models
Dr. Raquel Sousa
Advancing 3D drug development models from the in vitro to the ex vivo level
Conventional cellular or animal disease models have shown that the predictability of patient response to treatment with those models is severely limited. Great efforts have been made to humanize mouse models to better predict certain aspects of human physiology and immunology but with limited success. The abc biopply team has now made a significant breakthrough in humanizing upstream 3D cell models through the revolutionary and proprietary 3D CoSeedis multi-organoid in-chip communication technology™. Providing optimized physiological growth conditions and unique ways of intercellular communication, our models are freed from non-human components. Thus, they allow us to mimic and maintain physiologically relevant organoids in culture under reproducible and reliable conditions. Combined with the unique and statistically powerful predictiveness of the 3D CoSeedis™ chip, we are finally in a position to successfully bridge the translational gap between preclinical predictions and clinical treatments. Here we present how the innovative 3D CoSeedis in chip communication technology™ enables the humanization of 3D multi-organoid models and consequently improves the predictability of patient response. Preliminary data also suggests that the model is even capable of making accurate predictions on gender-specific drug responses.
LabTalk: From serum-free to xeno-free to defined media
Dr. Lothar Steeb
This lecture gives an overview of cell culture media, focusing on the progressive shift from serum-based formulations to serum-free alternatives, and subsequently to xeno-free and defined media systems. it will uncover the advantages and limitations associated with these media compositions, directly tested in our lab. Emphasizing the significance of defined media in modern cell culture practices, this lecture elucidates the pivotal role played by precise nutrient formulations in facilitating reproducibility, scalability, and regulatory compliance in diverse biological applications. Through a comprehensive review of current methodologies, innovations, and future prospects, this presentation aims to provide researchers and practitioners with insights essential for optimizing cell culture conditions and advancing biomedical research and bioprocessing industries.
Untargeted Metabolomics to Explore the Metabolic cues of Endothelial cells
Dr. Flávia Rezende
Metabolism research across all fields has recently and significantly advanced with the expansion of large-scale omics technologies. Unbiased methods became available to researchers at large covering the whole spectrum of elements within a cell or an organism. This enabled a better understanding of metabolic processes in a cell within a physiological context and from basic cell biology to biomedical and translational research. Our group focuses on the metabolic control of cardiovascular function and our aim is to find novel metabolites and metabolic pathways that modulate the function of endothelial cells. We have established assays and conditions for metabolomics and will share the tricks that allowed us to uncover a novel glutamine pathway that operates in endothelial cells and is important for their normal function.
Dr. Flávia Rezende is a biochemist and a junior group leader at the Department of Cardiovascular Physiology at the Goethe University in Frankfurt.
