Generation of a tumor spheroid in a microgravity environment as a 3D model of melanoma |
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Authors: | Bernadette Marrero Jane L Messina Richard Heller |
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Institution: | (1) Department of Molecular Medicine, University of South Florida, 12901 Bruce B. Downs Blvd., Tampa, FL 33612, USA;(2) Department of Pathology and Cell Biology, H. Lee Moffitt Cancer Center and Research Institute, University of South Florida, Tampa, FL, USA;(3) Frank Reidy Research Center for Bioelectrics, Old Dominion University, 830 Southampton Ave, Suite 5100, Norfolk, VA 23510, USA; |
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Abstract: | An in vitro 3D model was developed utilizing a synthetic microgravity environment to facilitate studying the cell interactions.
2D monolayer cell culture models have been successfully used to understand various cellular reactions that occur in vivo.
There are some limitations to the 2D model that are apparent when compared to cells grown in a 3D matrix. For example, some
proteins that are not expressed in a 2D model are found up-regulated in the 3D matrix. In this paper, we discuss techniques
used to develop the first known large, free-floating 3D tissue model used to establish tumor spheroids. The bioreactor system
known as the High Aspect Ratio Vessel (HARVs) was used to provide a microgravity environment. The HARVs promoted aggregation
of keratinocytes (HaCaT) that formed a construct that served as scaffolding for the growth of mouse melanoma. Although there
is an emphasis on building a 3D model with the proper extracellular matrix and stroma, we were able to develop a model that
excluded the use of matrigel. Immunohistochemistry and apoptosis assays provided evidence that this 3D model supports B16.F10
cell growth, proliferation, and synthesis of extracellular matrix. Immunofluorescence showed that melanoma cells interact
with one another displaying observable cellular morphological changes. The goal of engineering a 3D tissue model is to collect
new information about cancer development and develop new potential treatment regimens that can be translated to in vivo models
while reducing the use of laboratory animals. |
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