TY - JOUR
T1 - A novel stromal fibroblast-modulated 3d tumor spheroid model for studying tumor-stroma interaction and drug discovery
AU - Shao, Hongwei
AU - Moller, Mecker
AU - Wang, Dazhi
AU - Ting, Albert
AU - Boulina, Marcia
AU - Liu, Zhao Jun
N1 - Funding Information:
We thank Dr. Omaida C. Velazquez (University of Miami) for helpful collaboration, consultation, and discussion; Dr. Jie Li (University of Miami) for providing MeWo cells; and Dr. Meenhard Herlyn (The Wistar Institute) for providing all other melanoma cells. We also thank Dr. Marcia Boulina, Director of Analytical Imaging Core Facility, University of Miami, for imaging analysis. Zhao-Jun Liu was supported by grants from Bankhead-Coley Cancer Research Program (Award# 09BN-11), Women's Cancer Association (the 53rd annual grant) and internal funds from the University of Miami.
PY - 2020/2
Y1 - 2020/2
N2 - Tumor-stroma interactions play an important role in cancer progression. Three-dimensional (3D) tumor spheroid models are the most widely used in vitro model in the study of cancer stem/initiating cells, preclinical cancer research, and drug screening. The 3D spheroid models are superior to conventional tumor cell culture and reproduce some important characters of real solid tumors. However, conventional 3D tumor spheroids are made up exclusively of tumor cells. They lack the participation of tumor stromal cells and have insufficient extracellular matrix (ECM) deposition, thus only partially mimicking the in vivo conditions of tumor tissues. We established a new multicellular 3D spheroid model composed of tumor cells and stromal fibroblasts that better mimics the in vivo heterogeneous tumor microenvironment and its native desmoplasia. The formation of spheroids is strictly regulated by the tumor stromal fibroblasts and is determined by the activity of certain crucial intracellular signaling pathways (e.g., Notch signaling) in stromal fibroblasts. In this article, we present the techniques for coculture of tumor cells-stromal fibroblasts, time-lapse imaging to visualize cell-cell interactions, and confocal microscopy to display the 3D architectural features of the spheroids. We also show two examples of the practical application of this 3D spheroid model. This novel multicellular 3D spheroid model offers a useful platform for studying tumor-stroma interaction, elucidating how stromal fibroblasts regulate cancer stem/initiating cells, which determine tumor progression and aggressiveness, and exploring involvement of stromal reaction in cancer drug sensitivity and resistance. This platform can also be a pertinent in vitro model for drug discovery.
AB - Tumor-stroma interactions play an important role in cancer progression. Three-dimensional (3D) tumor spheroid models are the most widely used in vitro model in the study of cancer stem/initiating cells, preclinical cancer research, and drug screening. The 3D spheroid models are superior to conventional tumor cell culture and reproduce some important characters of real solid tumors. However, conventional 3D tumor spheroids are made up exclusively of tumor cells. They lack the participation of tumor stromal cells and have insufficient extracellular matrix (ECM) deposition, thus only partially mimicking the in vivo conditions of tumor tissues. We established a new multicellular 3D spheroid model composed of tumor cells and stromal fibroblasts that better mimics the in vivo heterogeneous tumor microenvironment and its native desmoplasia. The formation of spheroids is strictly regulated by the tumor stromal fibroblasts and is determined by the activity of certain crucial intracellular signaling pathways (e.g., Notch signaling) in stromal fibroblasts. In this article, we present the techniques for coculture of tumor cells-stromal fibroblasts, time-lapse imaging to visualize cell-cell interactions, and confocal microscopy to display the 3D architectural features of the spheroids. We also show two examples of the practical application of this 3D spheroid model. This novel multicellular 3D spheroid model offers a useful platform for studying tumor-stroma interaction, elucidating how stromal fibroblasts regulate cancer stem/initiating cells, which determine tumor progression and aggressiveness, and exploring involvement of stromal reaction in cancer drug sensitivity and resistance. This platform can also be a pertinent in vitro model for drug discovery.
KW - 3D tumor spheroid model
KW - Cancer Research
KW - Cancer-associated fibroblasts (CAF)
KW - Issue 156
KW - Melanoma initiating cells (MIC)
KW - Multicellular 3D spheroids
KW - Tumor microenvironment (TME)
KW - Tumor spheroids
KW - Tumor stem cells
KW - Tumor stromal fibroblasts
KW - Tumor-stromal interaction
UR - http://www.scopus.com/inward/record.url?scp=85081651380&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85081651380&partnerID=8YFLogxK
U2 - 10.3791/60660
DO - 10.3791/60660
M3 - Article
C2 - 32176195
AN - SCOPUS:85081651380
VL - 2020
JO - Journal of visualized experiments : JoVE
JF - Journal of visualized experiments : JoVE
SN - 1940-087X
IS - 156
M1 - e60660
ER -