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3-D Microwell Array System for Culturing Virus Infected Tumor Cells.
Rami El Assal1, Umut A Gurkan2,3, Pu Chen1
1Demirci 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.
Scientific Reports
|December 23, 2016
Summary
This study introduces a novel 3-D microwell system for culturing Kaposi
Area of Science:
- Oncology
- Virology
- Biotechnology
Background:
- Traditional 2-D cell cultures poorly mimic the tumor microenvironment.
- Accurate in vitro models are crucial for understanding cancer and developing therapies.
- Kaposi's sarcoma-associated herpesvirus (KSHV) infection requires advanced models for study.
Purpose of the Study:
- To develop an innovative 3-D cell culture model for KSHV-infected human B cells.
- To compare KSHV-infected cell behavior in 3-D versus 2-D cultures.
- To evaluate the potential of 3-D culture for oncology drug discovery.
Main Methods:
- Utilized a microwell array system for 3-D cell culture.
- Cultured KSHV-infected human B cells in 3-D for up to 15 days.
- Assessed viral genome copy number via immunofluorescence microscopy (LANA dots) and lytic reactivation rates.
Main Results:
- KSHV-infected B cells successfully cultured in 3-D for 15 days.
- 3-D cultures showed increased KSHV latency-associated nuclear antigen (LANA) dots, indicating higher viral DNA.
- Cells in 3-D exhibited a higher rate of lytic reactivation compared to 2-D cultures.
Conclusions:
- The 3-D microwell system effectively mimics the tumor microenvironment for KSHV-infected cells.
- This 3-D model enhances viral genome detection and lytic reactivation studies.
- The system offers a promising platform for improving 3-D oncology models and advancing drug discovery.
