TY - JOUR
T1 - Generation of homogenous three-dimensional pancreatic cancer cell spheroids using an improved hanging drop technique
AU - Ware, Matthew J.
AU - Colbert, Kevin
AU - Keshishian, Vazrik
AU - Ho, Jason
AU - Corr, Stuart J.
AU - Curley, Steven A.
AU - Godin, Biana
N1 - Funding Information:
B.G., M.J.W., S.J.C., K.C., and S.C. acknowledge financial support from NIH U54CA143837 and BG and MJW from NIH 1U54CA151668-01. The authors would also like to acknowledge Billie Smith and Sayeeduddin Mohammad from the Pathology and Histology core at Baylor College of Medicine, Houston, Texas.
Publisher Copyright:
© Mary Ann Liebert, Inc. 2016.
PY - 2016/4/1
Y1 - 2016/4/1
N2 - In vitro characterization of tumor cell biology or of potential anticancer drugs is usually performed using tumor cell lines cultured as a monolayer. However, it has been previously shown that three-dimensional (3D) organization of the tumor cells is important to provide insights on tumor biology and transport of therapeutics. Several methods to create 3D tumors in vitro have been proposed, with hanging drop technique being the most simple and, thus, most frequently used. However, in many cell lines this method has failed to form the desired 3D tumor structures. The aim of this study was to design and test an easy-to-use and highly reproducible modification of the hanging drop method for tumor sphere formation by adding methylcellulose polymer. Most pancreatic cancer cells do not form cohesive and manageable spheres when the original hanging drop method is used, thus we investigated these cell lines for our modified hanging drop method. The spheroids produced by this improved technique were analyzed by histology, light microscopy, immunohistochemistry, and scanning electron microscopy. Results show that using the proposed simple method; we were able to produce uniform spheroids for all five of the tested human pancreatic cancer cell lines; Panc-1, BxPC-3, Capan-1, MiaPaCa-2, and AsPC-1. We believe that this method can be used as a reliable and reproducible technique to make 3D cancer spheroids for use in tumor biology research and evaluation of therapeutic responses, and for the development of bio-artificial tissues.
AB - In vitro characterization of tumor cell biology or of potential anticancer drugs is usually performed using tumor cell lines cultured as a monolayer. However, it has been previously shown that three-dimensional (3D) organization of the tumor cells is important to provide insights on tumor biology and transport of therapeutics. Several methods to create 3D tumors in vitro have been proposed, with hanging drop technique being the most simple and, thus, most frequently used. However, in many cell lines this method has failed to form the desired 3D tumor structures. The aim of this study was to design and test an easy-to-use and highly reproducible modification of the hanging drop method for tumor sphere formation by adding methylcellulose polymer. Most pancreatic cancer cells do not form cohesive and manageable spheres when the original hanging drop method is used, thus we investigated these cell lines for our modified hanging drop method. The spheroids produced by this improved technique were analyzed by histology, light microscopy, immunohistochemistry, and scanning electron microscopy. Results show that using the proposed simple method; we were able to produce uniform spheroids for all five of the tested human pancreatic cancer cell lines; Panc-1, BxPC-3, Capan-1, MiaPaCa-2, and AsPC-1. We believe that this method can be used as a reliable and reproducible technique to make 3D cancer spheroids for use in tumor biology research and evaluation of therapeutic responses, and for the development of bio-artificial tissues.
UR - http://www.scopus.com/inward/record.url?scp=84963865395&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84963865395&partnerID=8YFLogxK
U2 - 10.1089/ten.tec.2015.0280
DO - 10.1089/ten.tec.2015.0280
M3 - Article
C2 - 26830354
AN - SCOPUS:84963865395
SN - 1937-3384
VL - 22
SP - 312
EP - 321
JO - Tissue Engineering - Part C: Methods
JF - Tissue Engineering - Part C: Methods
IS - 4
ER -