bims-orenst 2021-01-03 papers

Issue of 2021–01–03
four papers selected by
Joram Mooiweer, University of Groningen

Toxicology. 2020 Dec 23. pii: S0300-483X(20)30306-1. [Epub ahead of print] 152667

A 3D Microfluidic Liver Model for High Throughput Compound Toxicity Screening in the OrganoPlate®.

Kristin M Bircsak, Richard DeBiasio, Mark Miedel, Alaa Alsebahi, Ryan Reddinger, Anthony Saleh, Tongying Shun, Lawrence A Vernetti, Albert Gough.

We report the development, automation and validation of a 3D, microfluidic liver-on-a-chip for high throughput hepatotoxicity screening, the OrganoPlate LiverToxTM. The model is comprised of aggregates of induced pluripotent stem cell (iPSC)-derived hepatocytes (iHep) seeded in an extracellular matrix in the organ channel and co-cultured with endothelial cells and THP-1 monoblasts differentiated to macrophages seeded in the vascular channel of the 96 well Mimetas OrganoPlate 2-lane. A key component of high throughput screening is automation and we report a protocol to seed, dose, collect and replenish media and add assay reagents in the OrganoPlate 2-lane using a standard laboratory liquid handling robot. A combination of secretome measurements and image-based analysis was used to demonstrate stable 15 day cell viability, albumin and urea secretion. Over the same time-period, CYP3A4 activity increased and alpha-fetoprotein secretion decreased suggesting further maturation of the iHeps. Troglitazone, a clinical hepatotoxin, was chosen as a control compound for validation studies. Albumin, urea, hepatocyte nuclear size and viability staining provided Robust Z'factors > 0.2 in plates treated 72 hours with 180 μM troglitazone compared with a vehicle control. The viability assay provided the most robust statistic for a Robust Z' factor = 0.6. A small library of 159 compounds with known liver effects was added to the OrganoPlate LiverTox model for 72 hours at 50 μM and the Toxicological Prioritization scores were calculated. A follow up dose-response evaluation of select hits revealed the albumin assay to be the most sensitive in calculating TC50 values. This platform provides a robust, novel model which can be used for high throughput hepatotoxicity screening.

Keywords: hepatotoxicity; high throughput; iPSC-derived hepatocytes; liver-on-a-chip; screen; troglitazone

DOI: https://doi.org/10.1016/j.tox.2020.152667

J Neurochem. 2020 Dec 31.

Neuromuscular Junction-on-a-Chip: ALS disease modeling and read-out development in microfluidic devices.

Rianne de Jongh, Xandor M Spijkers, Svetlana Pasteuning-Vuhman, Paul Vulto, R Jeroen Pasterkamp.

Amyotrophic lateral sclerosis (ALS) is a fatal and progressive neurodegenerative disease affecting upper and lower motor neurons with no cure available. Clinical and animal studies reveal that the neuromuscular junction (NMJ), a synaptic connection between motor neurons and skeletal muscle fibers, is highly vulnerable in ALS and suggest that NMJ defects may occur at early stages of the disease. However, mechanistic insight into how NMJ dysfunction relates to the onset and progression of ALS is incomplete, which hampers therapy development. This is, in part, caused by a lack of robust in vitro models. The ability to combine microfluidic and induced pluripotent stem cell (iPSC) technologies has opened up new avenues for studying molecular and cellular ALS phenotypes in vitro. Microfluidic devices offer several advantages over traditional culture approaches when modeling the NMJ, such as the spatial separation of different cell types and increased control over the cellular microenvironment. Moreover, they are compatible with 3D cell culture, which enhances NMJ functionality and maturity. Here, we review how microfluidic technology is currently being employed to develop more reliable in vitro NMJ models. To validate and phenotype such models, various morphological and functional read-outs have been developed. We describe and discuss the relevance of these read-outs and specifically illustrate how these read-outs have enhanced our understanding of NMJ pathology in ALS. Finally, we share our view on potential future directions and challenges.

Keywords: ALS; iPSC; microfluidics; motor neurons; neuromuscular junction; organ-on-a-chip

DOI: https://doi.org/10.1111/jnc.15289

Biomaterials. 2020 Dec 21. pii: S0142-9612(20)30869-3. [Epub ahead of print]269 120622

Metastasis-on-a-chip reveals adipocyte-derived lipids trigger cancer cell migration via HIF-1α activation in cancer cells.

Jieun Seo, Kyeong Seog Kim, Jong-Wan Park, Joo-Youn Cho, Hak Chang, Junji Fukuda, Ki Yong Hong, Yang-Sook Chun.

Although obesity is a newly considered risk factor for cancer, the mechanisms by which adipocyte-derived metabolites accelerate cancer malignancy have yet to be elucidated. To identify the connection among heterogeneous cell types, conventional methods including Transwell assays or conditioned media (CM) have been used; however, these methods do not fully reflect niche effects in the tumor microenvironment (TME). Here, we established an oxygen permeable polydimethylsiloxane (PDMS)-based three-dimensional (3D) culture system to allow direct attachment between human adipocyte derived stem cells (ADSCs) and cancer cells. By doing so, a physiologically bioactive TME was created, which could be used to reveal further the relationships between different cell types. We found that co-culture of cancer cells with ADSCs resulted in a dispersion phenomenon, and the dispersed spheroid was well matched with the enhanced metastatic potential of cancer cells. Lipid profiling and in vitro migration assays suggested that lipids are the driving force for cancer cell migration via HIF-1α upregulation. In addition, the lipid/HIF-1α axis promoted tumor metastasis in a xenograft mouse model. This study presents an in vitro model of a biomimetic TME and provides new mechanistic insights into the effects of ADSC-released fatty acids on cancer cells as oncometabolites.

Keywords: 3D co-culture; ADSCs; Cancer cell migration; Fatty acids; HIF-1α; Tumor microenvironment

DOI: https://doi.org/10.1016/j.biomaterials.2020.120622

ACS Omega. 2020 Dec 22. 5(50): 32753-32760

Comprehensive Evaluation of Stable Neuronal Cell Adhesion and Culture on One-Step Modified Polydimethylsiloxane Using Functionalized Pluronic.

Wenming Liu, Meilin Sun, Kai Han, Rui Hu, Dan Liu, Jinyi Wang.

Polydimethylsiloxane (PDMS) is a popular and property-advantageous material for developing biomedical microsystems and advancing cell microengineering. The requirement of constructing a robust cell-adhesive PDMS interface drives the exploration of simple, straightforward, and applicable surface modification methods. Here, a comprehensive evaluation of highly stable neuronal cell adhesion and culture on the PDMS surface modified in one step using functionalized Pluronic is presented. According to multiple comparative tests, this modification is sufficiently verified to enable more significant cell adhesion and spreading in both quantity and stability, higher neuronal differentiation and viability/growth, more complete formation of the neuronal network, and stabler neuronal cell culture than the common coating tools on the PDMS substrate. The comparable and even superior cellular effects of this modification on PDMS to the standard coating of polystyrene for in vitro neurological research are demonstrated. Long-term microfluidic neuron culture with stable adhesion and high differentiation on the modified PDMS interface is accomplished, too. The achievement provides a detailed experimental demonstration of this simple and effective modification for strengthening neuronal cell culture on the PDMS substrate, which is useful for potential applications in the fields of neurobiology, neuron microengineering, and brain-on-a-chip.

DOI: https://doi.org/10.1021/acsomega.0c05190