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Breast cancer awareness

Development of a 3D Perfusion Platform for Breast Cancer Drug Testing

October 13, 2025 / by Lifeline® Cell Technology

National Breast Cancer Awareness Month

While huge strides have been made to increase breast cancer awareness nationwide, an estimated 1 in 8 women will be diagnosed with breast cancer in their lifetime. Initiatives promoting early detection through routine mammogram screening are contributing to improved patient outcomes, yet effective drug treatments remain limited, particularly for aggressive or treatment-resistant forms of the disease. In fact, industry reports found that oncology drugs had the lowest overall likelihood of approval (LOA) from Phase I among fourteen major disease areas.

Historically, researchers have relied on two-dimensional (2D) preclinical models for drug screening. While valuable, these models often fail to fully replicate the complex architecture and microenvironment of tumors in vivo, limiting their ability to predict clinical efficacy. To address these limitations, researchers are increasingly turning to three-dimensional (3D) tumor models, which more closely mimic the structure and behavior of real tumors. A recent study highlights the development of a perfused 3D culture system for preclinical testing of potential breast cancer therapeutics.

Validation of PerfusionPal, a 3D Cell Culture Model

Static 3D models are a significant improvement over traditional 2D cultures; however, many still fail to capture the dynamic biochemical and biophysical cues provided by fluid flow, which can critically influence cancer cell behavior. In this study, Liu and colleagues validated the PerfusionPal perfused culture system as a fully humanized method for breast cancer cell culture. The platform was evaluated using patient samples from ER-positive breast cancer and established breast cancer (BC) cell lines to compare tumor growth and tamoxifen (TAM) response under static versus perfused 3D conditions.

The PerfusionPal system consists of an integrated multi-well insert containing SeedEZ scaffolds—clear, hydrophilic disks of randomly arranged glass fibers. BC cell lines or primary explants were seeded onto SeedEZ disks and cultured under either static or perfused conditions. Perfusion was achieved using a proprietary bidirectional in-well perfusion method, where a high-density, breathable hemoglobin analogue (“Blood Substitute”) was delivered via syringe pump to mimic oxygen delivery in blood.

All BC lines (MCF-7, BT-474, and MDA-MB-231) were cultured in Lifeline® Cell Technology’s FibroLife® Serum Free Medium, a formulation free of animal-derived components and further modified by omitting FGF and rhEGF/TGF-β (referred to as epiFL), before establishing 3D cultures. Across both static and perfused cultures, parameters including metabolic activity (AlamarBlue assay), proliferation, and response to TAM (cell viability by fluorescence intensity) were measured.

Perfusion improved the homeostasis of the 3D culture environment and significantly enhanced metabolic activity and proliferation, particularly during long-term culture (up to 21 days). TAM responses were initially similar between static and perfused models; however, after day seven, perfused cultures displayed greater drug sensitivity, especially at higher TAM doses. At 100 nM TAM, perfusion resulted in a significantly stronger inhibitory effect on cell viability by day fourteen compared to static culture.

Similarly, patient-derived samples cultured under perfusion demonstrated improved metabolic activity and enhanced responsiveness to TAM. These findings are encouraging, as establishing patient-derived xenograft (PDX) models from ER-positive BC is notoriously challenging, with reported engraftment rates below 15%. PerfusionPal’s ability to sustain patient tissue in vitro suggests it could offer a more accessible and reliable platform for evaluating therapeutic responses.

Importantly, PerfusionPal distinguishes itself from microfluidic devices, which often expose cells to damaging shear stress and are prone to clogging, which is problematic for long-term culture. By enabling culture at a macro scale, PerfusionPal allows use of larger amounts of native patient tumor tissue, helping preserve the intratumoral genetic and cellular heterogeneity, preserving the in vivo tumor microenvironment.

Based on the data, the authors conclude that the fully humanized PerfusionPal platform holds as a tool for preclinical drug testing and personalized medicine applications while also overcoming some of the ethical and scientific disadvantages associated with animal models in breast cancer research.

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Lifeline Cell Technology offers a comprehensive portfolio of reproductive and mammary cells (male and female), in addition to specialized media that are optimized for their growth to help researchers answer important research questions:

Join us next month for another installment of the Lifeline® blog to see how our cells and culture media are advancing biomedical research worldwide. If you have used our products in your publication, we’d love to feature your work here!

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