Poster B1 - A low-cost, high-throughput engineered breast tumor model for drug testing

Thursday, October 24, 2024

10:00 AM – 11:00 AM EST

Location: Exhibit Hall E, F & G

Co-Author(s)

Elvan Dogan Kumtepe, Dr.

PhD, research assistant
New Jersey institute of technology
West Orange, New Jersey, United States

Presenting Author(s)

Vidhi Patel

Student
New Jersey Institute of Technology, United States

Primary Investigator(s)

Amir K. Miri, PhD (he/him/his)

Assistant Professor
New Jersey Institute of Technology
Newark, New Jersey, United States

Introduction: High mortality rates in breast cancer patients are primarily due to metastatic tumor spread, recurrence, and resistance to clinical treatments. The mechanisms behind drug-resistant tumors and their recurrences remain poorly understood. The aggressive nature of metastatic cancer highlights the urgent need for more precise treatments. Patient-derived xenograft models have been developed to identify the most effective drugs for individual patients. These models suffer from limitations such as low success rates, excessive cost of manufacturing, and time-consuming processes, which reduce their effectiveness in drug screening. To advance our understanding of tumor cell mechanisms, there is a need for engineered models that replicate the tumor cell response within a mimetic tumor microenvironment (TME). Advanced Biofabricaion Lab (NJIT) has developed a hydrogel-based microfluidic system for 3D tumor modelling using a gelatin-based matrix with tunable mechanical properties [1, 2]. This model is designed to create a cost-effective, high-throughput hydrogel-based screening platform for personalized medicine. Our TME model simplifies the testing of various drugs and their effects on tumors. It conserves resources and allows for the reuse of portions of the chip for multiple tests, using laser cutting, hydrogel engineering, and precise biologics positioning. This research introduces an efficient method for drug testing with minimal resource usage.

Materials and

Methods: We purchased all materials and assays from Thermo Fisher and Sigma Aldrich. Spheroids of MDA-MB-231 cells (ATCC) and NHLF (ATCC), mixed in a 1:1 ratio, were formed at a density of 15,000 cells per well in ultra-low attachment 96-well plates over a period of three days. Gelatin methacryloyl (GelMA) was synthesized, and a 10% w/v GelMA precursor was prepared according to a well-established protocol [2]. A high-throughput perusable microfluidic chip was designed and fabricated using acrylic sheets, double-sided tape, and a laser cutter. The optimal laser cutter settings were optimized in terms of power and speed for the etching portion of the chip (depth of 500 um) and making cuts. Precise positioning of the spheroids was achieved through negative pressure with a 30-gauge needle. A resin printer was used to print the tubing adapter to create leak free chip. Experimental studies were conducted to determine the optimal vacuum limits for spheroid positioning, as part of our related project on precise spheroid placement. After positioning and crosslinking, the chip was supplied with optimized cell culture media using an applied using syringe pump and after washing, fluorescent images are obtained for viability quantification.

Results, Conclusions, and Discussions: The spheroids after being placed in the wells of the chip with GelMA, they are incubated with active perfusion (Figure 1). To monitor the survival, images and size of the spheroids were taken daily, which show a growth in size. Tumor growth and viability within a controlled microenvironment was investigated across samples. Spheroids were observed to gradually migrate within the hydrogel microfluidic chip under active perfusion, indicating the effectiveness of the designed setup in facilitating controlled TME. Following the incubation period of 10 days, there were no leakages present as well as the fluorescent images revealed a high survival rate of the co-cultured tumor spheroids. The success of this chip leaves room to create a cost and time effective method for testing cancer medication especially as the materials are commonly used lab materials. Low cost of construction for testing allows for medication Doxorubicin to test personalized treatment plans for patients. Along with the efficiency, the method that we optimized can be adapted to test for multiple types of cancer by optimizing the GelMA bioink. The success of the chip creates an accurate 3D model to perform personalized method of testing medication. Each part of the structure mimics what the spheroids would experience in the body and therefore provides data that is physiologically relevant.

Conclusion: Future work for this chip is creating a concentration gradient to test medication. We optimized a method of testing that can accurately predict the effectiveness of medication to provide patients with a personalized treatment plant. This method works by using lab materials to make it low cost and mimic the environment of the body.

Acknowledgements (Optional):