Having a human cell-based model that mimics the in vivo tumor microenvironment and cell-to-cell interactions is invaluable for studying new cancer therapies. In our last post, we talked about growing spheroids using our idenTx family of 3D tissue culture chips and plates to reproduce in vivo data related to the use of PD-1 blockade during adoptive T cell therapy. In this article, we’re going to look in more detail just how those models are created from mouse or patient biopsies, and some of the possible output data that can be obtained using idenTx in combination with standard lab equipment.
Obtaining high quality spheroids
In 2018, Aref and collaborators published a study demonstrating step-by-step how to create 3D in vitro cancer models using AIM Biotech’s idenTx chips.
This work highlights the importance of starting with quality biopsies—meaning fresh samples with high cellular viability. The scientists produced tissue fragments from these biopsies that were later mixed with collagen and injected into the idenTx chips. They maintained these spheroids in the 3D cell culture device for 5 to 9 days while performing the desired experiments. During that time, they applied anti-PD-1 antibodies to simulate PD-1 blockade therapy.
Readouts obtained from idenTx chip experiments
idenTx: experimental flexibility and a range of meaningful analyses
Microscopy
During the injection of spheroids to the chip, scientists estimated the number of injected cells using phase-contrast microscopy, estimating about 10,000 to 20,000 cells per microfluidic chamber. They also used live (time-lapse) microscopy to track spheroid formation and behavior within
the idenTx chip over time. Finally, to measure cell death relative to anti-PD-1 treatment efficacy, the authors utilized immunofluorescence staining by injecting the dye directly into the idenTx chip.
Media collection and analysis
As the scientists were interested in the early immune response to anti-PD-1 treatment, they collected the cell culture media from the chip and used it to run assays that quantify the concentration of different cytokines and chemokines on the collected sample.
Cell collection and analysis
Additionally, the authors analyzed the RNA levels from the tissue treated with anti-PD-1 to determine whether the treatment changed the immune cell populations. For that, they injected the cell lysis reagent into the idenTx chip in order to later collect the sample, purify the RNA, and perform the sequencing.
This study highlights how AIM’s researcher-friendly idenTx chips work together with standard lab tools—no need for special equipment to employ any of the techniques laid out in this paper, unlike many competing organ-on-a-chip platforms. Experimental manipulations, human-relevant data collection, and downstream analysis were all done without need to purchase or learn any additional appliances—just AIM’s elegant organ-on-a-chip assays, and same research instruments used for any other such test.
Gathering more predictive, human-relevant data is what AIM Biotech is all about. Want to discuss how idenTx can transform your drug discovery research? Use the chat bubble on the bottom right corner of this page, and we’ll reach out to you—or check out our Contact Us page. Also be sure to look at how our contract research services can help streamline your workflow.
*3D microfluidic ex vivo culture of organotypic tumor spheroids to model immune checkpoint blockade. Aref AR et al. Lab Chip. 2018 Oct 9;18(20):3129-3143. doi: 10.1039/c8lc00322j. PMID: 30183789; PMCID: PMC6274590.