Organoids: Exploring Liver Cancer Initiation and the Possibilities of Personalized Glioblastoma Treatment – Technology Networks

In the search for improved and high-throughput in vitro models, organoids have emerged as a promising 3D cell culture technology.1 Defined as a three-dimensional multicellular in vitro tissue construct, organoids are derived from cells that spontaneously self-organize into properly differentiated functional cell types to mimic at least some function of an organ.2 Organoid formation is driven by signaling cues in the extracellular matrix and medium, and is influenced by the particular cell types that are present.2 Compared with two-dimensional cultures, organoids incorporate more physiologically relevant cell-cell and cell-matrix interactions, and are a better reflection of the complex network found in vivo.With significant opportunities for studies of human-specific disease mechanisms, personalized medicine, drug discovery, pharmacokinetic profiling and regenerative medicine, organoids are being pursued across a range of disciplines. Many anticipate that these cell culture models will result in more efficient translation of research into clinical success. In this article, we explore the various types of organoids under development and shine a spotlight on some of the different approaches to organoids in cancer research.

Organoids can be derived from pluripotent stem cells (including embryonic stem cells or induced pluripotent stem cells) or neonatal or adult stem cells from healthy or diseased tissue.1,2 Cancer organoids have been generated from a range of human cancer tissues and cell lines including colon, pancreas, prostate, liver, breast, bladder and lung.6-12 This year, a research group led by Hongjun Song, Professor of Neuroscience at the Perelman School of Medicine at the University of Pennsylvania, published a report in Cell detailing methods for the rapid generation of patient-derived glioblastoma organoids.13Fresh tumor specimens were removed from 53 patient cases to produce microdissected tumor pieces that could survive, develop a spherical morphology and continuously grow in culture for at least two weeks (Figure 1). The production of glioblastoma organoids was achieved while maintaining a high level of similarity between the organoids and their parental tumors, with the expression levels of specific markers showing stability over long-term culture (48 weeks). Importantly, native cell-cell interactions were preserved by avoiding mechanical and enzymatic single-cell dissociation of the resected tumor. As Song explains, this was achieved on a clinically relevant timescale: Normally, the treatment for glioblastoma patients starts one month after surgery. The idea is that glioblastoma organoids can be generated within two weeks and subjected to testing of different treatment strategies to come up with the best option for a personalized treatment strategy.

Figure 1: Glioblastoma organoid generation, from fresh tumor pieces to frozen spherical organoids. Image used with permission from Jacob et al. 2020.One concern with organoid formation and expansion is the potential variability of the serum or Matrigel that can exist across batches and sources, creating variable exogenous factors that could cause the organoid to divert. This ultimately compromises reproducibility, a major bottleneck of current organoid systems.2,13 To avoid this source of error, Songs group used an optimized and defined medium devoid of variable factors that could contribute to the clonal selection of specific cell populations in culture.Glioblastoma is the most prevalent primary malignant brain tumor in adults,14 and having glioblastoma organoids available for research would present significant opportunities, explains Song: They can be used to test different drugs based on mutation profiles and to investigate mechanisms underlying tumor progression, drug sensitivity and resistance. While the accuracy of these predictions would need to be verified, researchers hope that patient-derived organoids will be used to help inform oncologists, accelerate drug discovery, and lead to better clinical trial design.Live-Cell Monitoring: Optimizing Workflows for Advanced Cell Models

As cell-based assays become technically more complex, the need to holistically capture dynamic and sometimes subtle cellular events becomes ever more important. By providing real-time imaging data of cellular events without disturbing the sample during the cell culture workflow, live-cell monitoring can support the optimization of these advanced models. Download this whitepaper to discover how live-cell monitoring can support such optimization, with a breadth of applications.