Leukaemia is a form of cancer that affects cells of the haematopoietic lineage. Acute myeloid leukaemia (AML) and chronic lymphocytic leukaemia (CLL) are the two most common types, accounting for 31% and 38% of all leukaemia diagnoses in the United Kingdom.
The study of leukaemia has been traditionally conducted in two-dimensional (2D) cultures or resorting to animal models, both systems failing to recapitulate the physiological parameters of the disease. A three-dimensional (3D) bone marrow model comprised of polyurethane (PU) scaffolds coated with Collagen I has been shown to allow the culture of leukaemic cell lines, showing potential towards the culture of primary cells. In this thesis, this 3D model was tailored to the culture of primary bone marrow (BM) and peripheral blood (PB) cells of patients with leukaemia, using serum-free cytokine-free conditions and enabling a long-term culture (>28 days). Scaffold dimensions, geometry, cell seeding methods, extraction methods, coating proteins and storage protocols were studied and optimised.
The study of AML in this platform allowed the xenofree culture of PB and BM cells for up to 6 weeks, maintaining >80% viability, 30 days longer than the previously reported primary AML culture. The long-term viable cultures allowed the study of population dynamics through flow cytometry, measurement of cell expansion and delivery of chemotherapy, highlighting higher susceptibility of primary cells to drugs. The study of CLL allowed for long-term cultures of up to 8 weeks of cells derived from PB and BM, demonstrating that 3D and serum-free conditions reduced CLL cell apoptosis in vitro, allowing the delivery of a novel BCL2 inhibitor. This is the first long-term serum-free, cytokine-free and feeder-cell free culture to be reported.
In summary, this thesis reports the long-term xenofree study of primary leukaemia in a PU scaffold, allowing the study of population dynamics and drug delivery.