Design and validation of a bone marrow-on-chip platform for cancer modeling and therapy testing

The bone marrow is a highly specialized microenvironment that plays a central role in both physiological hematopoiesis, as well as pathological processes including the progression of hematological malignancies. A major limitation in current cancer research is the lack of physiologically relevant in vitro models capable of reproducing the dynamic cellular and molecular interactions occurring within the human bone marrow niche. This thesis presents the design, fabrication, and preliminary biological validation of a two-layer PDMS mesofluidic bone marrow-on-chip platform intended as a versatile tool for cancer modeling and therapeutic testing. The device comprises a lower compartment containing a Gelatin Methacryloyl (GelMA) hydrogel and an upper fluidic channel. Plasma bonding is employed to assemble the two PDMS layers, enabling controlled biochemical communication between the upper channel and the underlying hydrogel compartment. In the initial validation phase, the platform is engineered to recapitulate key features of the physiological bone marrow stromal niche. The lower hydrogel compartment is populated with human mesenchymal stem cells (hMSCs) and BJ, while the upper fluidic channel is lined with human umbilical vein endothelial cells (HUVECs). This configuration successfully established a vascularized interface and demonstrated the device's ability to support complex stromal-endothelial interactions in a 3D environment. Building upon this validation, the platform was subsequently adapted to model a leukemic microenvironment by embedding human acute myeloid leukemia (AML) cells within the GelMA hydrogel. In this pathological configuration, the upper fluidic channel is designed to allow the controlled perfusion of patient-derived monocytes previously infected ex vivo with an oncolytic herpes simplex virus type-1 (oHSV1). This setup is intended for future studies investigating monocyte-mediated viral delivery and oncolytic activity against leukemic cells under three-dimensional culture closely mimicking the in vivo conditions. Overall, this work establishes a mesofluidic bone marrow-on-chip model as a controllable and physiologically relevant in vitro system. It demonstrates the successful reconstruction of the stromal-vascular niche and highlights the platform potential for investigating tumor-microenvironment interactions and assessing emerging cell- and virus-based anticancer therapies.