UBC Theses and Dissertations
Development and application of an in vitro culture model for primary human T-ALL cells Yost, Arla June
T-cell acute lymphoblastic leukemia (T-ALL) is an aggressive hematologic malignancy that affects both children and adults. Optimization of chemotherapy regimens has led to steady improvements in outcome for pediatric patients over the last 5 decades, with a long-term survival rate of 80%. However, the five-year survival rate for adults is still only 35-40% and there is a poor prognosis for relapse patients (Goldstone et al., 2008). Further improvements in outcome will undoubtedly require introduction of novel approaches and more specific targeted therapies. Research efforts in this area have been hampered by the lack of a reproducible model for in vitro growth of human T-ALL blasts. Most efforts to date have relied heavily upon established cell lines, which can be a useful tool to study malignancies but do not necessarily always represent bona fide disease biology, and in vivo studies involving patient samples expanded as xenografts in immunocompromised mice, which are costly, time-consuming and complicated by non-cell autonomous effects. Current methods for in vitro culture of patient T-ALL samples yield variable performance with high rates of apoptosis and less than robust proliferation. Development of an efficient and reproducible in vitro culture method for growth of primary human T-ALL blasts would greatly enhance the ability to test and validate novel therapies by allowing for direct assay for sensitivity/resistance of patient cells which have not been subject to extensive manipulation or selection. In this work we report an in vitro co-culture system using defined, serum-free media and a stromal feeder cell layer which supports robust growth and minimal apoptosis of patient T-ALL blasts. We have shown that the stromal feeder cell layer and supplemental IL-7 cytokine is critical for sustained patient T-ALL blast growth in this model. Finally, we have demonstrated the utility of this culture system as a platform that will facilitate ongoing efforts to identify growth factors/cytokines required for maintenance of leukemia cell self-renewal activity, aid in the study of signaling pathways important in T-ALL pathogenesis and maintenance, and allow for prospective testing of novel compounds for therapeutic efficacy on patients’ own tumor cells, thus enabling implementation of personalized medicine initiatives.
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