PELONEWS Autumn 2023

Overview Organoids

Overview Organoids PELONews Autumn 2023 Reprinted from “Organoid Generation from Pluripotent Stem Cells”, by BioRender.com (2023). Organoids are three-dimensional miniaturized organ-like structures that are derived from stem cells or tissue samples. They are composed of multiple cell types and closely mimic the structure and function of specific organs or tissues in the body. Organoids offer a unique platform for studying organ development, disease modelling, and personalized medicine. They provide a more accurate representation of human biology compared to traditional two-dimensional cell cultures, allowing researchers to investigate complex cellular interactions and responses to various stimuli. Organoids hold great promise in advancing our understanding of organ physiology, drug discovery, and regenerative medicine. PELONews: Organoids, Unlocking the power of 3D Tissue Models for Disease Research and Drug Discovery Scan/Click the QR code to read our article to find out. Why is defined media better for organoids? • Ethics: FBS is derived from the blood of unborn calves, raising ethical concerns regarding animal welfare. In contrast, defined media eliminate the need for FBS, reducing reliance on animalderived components. . • Consistency: Defined media ensures consistent and reproducible manufacturing as it is precisely formulated. • Control: Precisely formulated composition provides control over exosome production and thereby improves scalability and efficiency in manufacturing • Optimization: Tailoring media composition enhances exosome yield, purity, and functionality. • Standardization: Eliminating undefined components like serum ensures standardized exosome preparation and uniform reproducible results. • Safety: Minimizing contaminants and unwanted effects with defined media ensures high-quality exosomes by eliminating adverse immune reactions associated with undefined components.. • Compliance: Regulatory guidelines and ethical considerations are met in exosome production. Defined media reduce the risk of potential contaminants or unwanted biological effects. By precisely defining the media composition, unwanted variations and impurities can be minimized, ensuring the production of high-quality organoids. 2 I www.pelobiotech.com

Overview Organoids Table 1. Comparison of Cell Culture Factors for Cell Growth and Exosome Studies. Adapted from Liau et al, Int J Mol Sci, 2021 Factors FBS hPL Defined Media EXO Free Media Pros Abundant and accessible Easy to produce Well-defined components Exosome-depleted and ideal for isolation Suitable for most cells Suitable for most cells No batch-to-batch variation No batch-to-batch variation Supports cell survival and proliferation Supports cell survival and proliferation No risk of xenogeneic/ allogeneic protein contamination No risk of xenogeneic/allogeneic protein contamination Widely used in studies and publications No risk of xenogenic immune reaction; can be produced using autologous blood No unintended interaction with test substances No unintended interaction with test substances Cons Components are illdefined Components are illdefined Not available for certain cells Not available for certain cells Batch-to-batch variation Batch-to-batch variation (reduced by pooling) Some media require extra coating for cell attachment Some media require extra coating for cell attachment Potential contamination (prions, mycoplasma, bacteria, fungi) Unintended interaction with test substances More expensive compared to alternatives More expensive compared to alternatives Risk of animal protein contamination in clinical usage Few distributors Time-consuming and difficult to develop Time-consuming and difficult to develop Unintended interaction with test substances Potential allogenic immune response Higher particle count and presence of exosomes Ethical concerns with animal welfare Risk of transmitting human viruses Multitude of exosomes not suitable for isolation or studies Multitude of exosomes not suitable for isolation or studies 3