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Cancer cells have been increasingly grown in pharmaceutical research to understand tumorigenesis and develop new therapeutic drugs. Currently, cells are typically grown using two-dimensional (2-D) cell culture approaches, where the native tumor microenvironment is difficult to recapitulate. Thus, one of the main obstacles in oncology is the lack of proper infection models that recount main features present in tumors. In recent years, microtechnology-based platforms have been employed to generate three-dimensional (3-D) models that better mimic the native microenvironment in cell culture. Here, we present an innovative approach to culture Kaposi’s sarcoma-associated herpesvirus (KSHV) infected human B cells in 3-D using a microwell array system. The results demonstrate that the KSHV-infected B cells can be grown up to 15 days in a 3-D culture. Compared with 2-D, cells grown in 3-D had increased numbers of KSHV latency-associated nuclear antigen (LANA) dots, as detected by immunofluorescence microscopy, indicating a higher viral genome copy number. Cells in 3-D also demonstrated a higher rate of lytic reactivation. The 3-D microwell array system has the potential to improve 3-D cell oncology models and allow for better-controlled studies for drug discovery.
NSF Career 1461602; NIH R01DE02497101; NIH R21AI110277; NIH R21AI113117; NIH R01 AI093282; NIH R01DE024971; NIH CA082036; NIH R01 GM108584; NIH R01 AI120683; NIH R01 AI122862; NIH U54EB015408; NIDCR DE025208
National Science Foundation (NSF); National Institutes of Health (NIH)
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Gurkan, Umut A., "3-D Microwell Array System for Culturing Virus Infected Tumor Cells" (2016). Faculty Scholarship. 29.