We are trying a new event at the annual meeting “Tech in C-TEC”, a showcase of technology and innovation from our community. The aim of this event is to provide an opportunity for members to demonstrate new technology to the group to facilitate technology sharing and catalyse future collaborations. Come along to learn about innovations in computer and laboratory techniques from your colleagues in Cancer TEC.
A microfluidic rheometer for tumor mechanics
Mingming Wu, Cornell University
YoungJoon Suh1, Mrinal Pandey1, Alan Li 1, Herbert Hui2, BangGuo Zhou2, Jeffrey E Segall3, and Mingming Wu1* 1 Biological and Environmental Engineering Department, 2Mechanical and Aerospace Engineering Department, Cornell University, Ithaca, NY. USA. 3 Department of Pathology, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, NY. USA.
Clinically, the feel, touch, and shape of a solid tumor are important diagnostic methods for malignant state of a tumor. However, there are limited tools for quantifying the mechanics of the tumor, and its relevancy to the malignant state. In this presentation, I will showcase a microfluidic rheometer recently developed in my lab. This device, size of your palm, allows for characterizing mechanics of tumor spheroids while the spheroids are embedded within a 3D matrix. The enabling feature of the device is its ability to do living material characterization in a physiologically realistic 3D setting. I will highlight the details of how the device is made, and how it works.
A Novel Tissue Bioreactor for Retinal Organoid Microenvironmental Control
Emma Drabbe, University of Miami
In vitro culture systems generally apply homogeneous stimuli and rely on intercellular signaling to guide growth of tissues. However, to derive complex tissue structures such as the human retina, a gradation of certain stimuli is required. The inner retina resides in a hypoxic environment (2% O2) adjacent to the vitreous cavity. From there, oxygenation levels rapidly increase towards the outer retina (18% O2) at the choroid. Here we developed a novel tissue bioreactor allowing the maturation of inner and outer retinal cell phenotypes within an O2 gradient. The bioreactor is assembled from an acrylic slide, a gas-permeable film, a cover glass, and double-sided adhesives, which were adjusted with computer numerical control milling and laser cutting. The 55 culture wells of 1.5 mm in diameter and 0.7 mm high each hold one retinal organoid. A sodium sulfite solution provides the bioreactor with an oxygen leaching channel and a dual syringe pump creates a 50 µL/hr continuous flow of culture medium though the bioreactor. The gas diffusion throughout the culture medium resulted in an O2 concentration gradient along the z-axis. The computational predictions in atmospheric conditions are in accordance with measurements around the retinal organoid location in the bioreactor. This open-well bioreactor is easily accessible for downstream analysis, establishes a steep O2 gradient and allows high-throughput retinal organoid culture. It will help retinal organoids mature into the complex structure to use them for disease modeling and drug testing of Retinoblastoma.
HypoxyCaps for Subwell Control of Oxygen Levels
Kris Killian, UNSW Sydney, and Tom Molley, UCSD
Hypoxia plays a critical role in tumor progression and metastasis, therapeutic resistance, and cancer biology. However, current tools to replicate physiological oxygen conditions in the laboratory are costly, complex, and inaccessible for many researchers. These challenges hinder the development of accurate preclinical models essential for advancing cancer therapies. To address this unmet need, Drs. Molley and Kilian present HypoxyCaps, a novel technology designed to revolutionize in vitro modeling of hypoxia for cancer research. HypoxyCaps are enzyme-loaded microcapsules that reduce oxygen levels in cell culture media with unparalleled precision and simplicity. This innovative platform allows researchers to generate hypoxic conditions in individual wells, create oxygen gradients, and establish multiple oxygen levels across well plates—without the need for specialized equipment. The technology is versatile, cost-effective, and adaptable for both high-throughput cancer studies and advanced microfluidic models. This interactive demonstration will feature live experiments showcasing HypoxyCaps in action. Attendees will observe real-time oxygen reduction using colorimetric readouts and live oxygen measurements in well plate and microfluidic chip formats. Additionally, hands-on engagement will allow participants to explore the physical properties of the capsules and their ease of use. By lowering the barriers to studying hypoxia in vitro, HypoxyCaps enable researchers to create more physiologically relevant models of tumor microenvironments. This advancement has the potential to accelerate discoveries in cancer biology and therapeutic development, ultimately bridging the gap between laboratory research and clinical application.
In-Plane and Out of Plane Tissue Stretcher withOptical Capabilities
Luis Solorio, Purdue University
Current in vitro 3D cell culture platforms used to study cell response to mechanical stimuli require seeding cells on synthetic materials, thereby failing to capture the effect of extracellular matrix proteins on cellular adhesion and signaling pathways. Further, characterizing material response to mechanical stimulation while imaging biological materials remains a challenging endeavor for the field of tissue engineering. Here, we developed a simple, cost-effective, uni-axial stretching platform with high-resolution force sensing capabilities tailored for microscopy cell culture experiments. The platform can apply a constant tensile load, or a cyclic load depending on the dynamics of the physiological condition being modelled.
Microphysiological systems capturing the impact of ovulation on early events in ovarian cancer
Jonathan Coppeta, Charles Stark Draper Laboratory
Ovarian cancer remains the 5th leading cancer death in women with only 50% survival beyond 5 years. Of the ovarian cancer sub-types, high-grade serous carcinoma (HGSC) accounts for almost 70% of all tumors and has the highest mortality. The cell of origin for HGSC was previously thought to be from the ovarian surface, but it is now clear that the fallopian tube epithelium is the likely source of most HGSC and that the ovary is a primary metastatic site. A major clinical issue remains the lack of information with regards to early disease formation that can be integrated into new biomarkers, prevention, and treatment strategies. To address this unmet need, we have developed the PREDICT-MOS microfluidic tissue culture system which supports key aspects of early disease modeling including, i.) precision tissue perfusion which has been shown to maintain ovarian and fallopian tissue function over extended culture durations (1 month), ii.) programmable ovarian-fallopian microfluidic interactions, iii.) facile access to tissues for nucleic acid analysis and secreted factors (e.g., extracellular vesicles, metabolites, and secreted proteins), iv.) supports cellular migration and invasion, and v.) supports multiple 2D and 3D tissue formats. The PREDICT-MOS system has supported long-term ovary-fallopian interactions over a full menstrual cycle and we currently have published 14 papers that cover aspects of this biology from the role of key tumor suppressors, the development of preneoplastic lesion models, the expansion of stem cell populations, and the role of secreted ovarian factors as homing signals for fallopian tube cell expansion within the ovary.
Brian P. Cain2, Jane Miglo1, Kathy De La Torre1, Angela Russo1, Brett C. Isenberg2, Joanna E. Burdette1, Jonathan R. Coppeta2 1Department of Pharmaceutical Sciences, University of Illinois Chicago College of Pharmacy, Chicago, IL, United States. 2Charles Stark Draper Laboratory, Cambridge, MA, United States.
OrganixInsights: High content profiling of Organoid Models
Scott Sax, University Nevada Reno
Profiling of Patient-Derived organoids is necessary for drug screening and precision medicine. This task requires an integrated Imaging Bioinformatics system that integrates experimental variables with image-based data and a computational engine to process 3D images. OrganixInsights is designed to meet these requirements. The informatics component provides a layer for multifactorial experimental design, visualization of 3D images, and meta-analysis. The computational component provides accurate segmentation of three-dimensional cellular organization followed by protein readouts. While fully Convolutional Neural Networks are widely used in medical image segmentation, they struggle to capture long-range dependencies necessary for accurate segmentation. On the other hand, transformer models have shown promise in capturing long-range information across domain boundaries. Motivated by this, 3D-Organoid-SwinNet provides the computational engine for processing large amounts of organoid samples that are imaged in 3D with a minimal number of parameters.