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Sci Rep. 2016 Dec 22;6:39144. doi: 10.1038/srep39144.

3-D Microwell Array System for Culturing Virus Infected Tumor Cells.

Author information

1
Demirci Bio-Acoustic-MEMS in Medicine (BAMM) Laboratories, Canary Center at Stanford for Cancer Early Detection, Department of Radiology, Stanford University School of Medicine, Palo Alto, CA, 94304, USA.
2
Demirci Bio-Acoustic-MEMS in Medicine (BAMM) Laboratories, Division for Biomedical Engineering, Division of Infectious Diseases, Renal Division, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, 02115, USA.
3
Biomanufacturing and Microfabrication Laboratory, Mechanical and Aerospace Engineering Department, Biomedical Engineering Department, Department of Orthopedics, Advanced Platform Technology Center, Louis Stokes Cleveland Veterans Affairs Medical Center, Case Western Reserve University, Cleveland, OH, 44106, USA.
4
Division of Infectious Diseases, Departments of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, 02115, USA.
5
Department of Electrical Engineering (by courtesy), Stanford University School of Engineering, Stanford, CA, 94305, USA.

Abstract

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.

PMID:
28004818
PMCID:
PMC5177905
DOI:
10.1038/srep39144
[Indexed for MEDLINE]
Free PMC Article

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