TY - JOUR
T1 - Generation and Expansion of Primary, Malignant Pleural Mesothelioma Tumor Lines
AU - Griffiths, Tamara M.
AU - Ramos, Carlos
AU - Haymaker, Cara L.
N1 - Funding Information:
We would like to acknowledge Raquel Laza-Briviesca for her contribution to starting this protocol and Drs. Boris Sepesi, Reza Mehran, and David Rice for collaboration on tissue collections. There is no additional funding associated with this work.
Publisher Copyright:
© 2022, Journal of Visualized Experiments. All rights reserved.
PY - 2022/4
Y1 - 2022/4
N2 - Current methodologies for the expansion of primary tumor cell lines from rare tumor types are lacking. This protocol describes methods to expand primary tumor cells from surgically resected, malignant pleural mesothelioma (MPM) by providing a complete overview of the process from digestion to enrichment, expansion, cryopreservation, and phenotypic characterization. In addition, this protocol introduces concepts for tumor generation that may be useful for multiple tumor types such as differential trypsinization and the impact of dissociation methods on the detection of cell surface markers for phenotypic characterization. The major limitation of this study is the selection of tumor cells that will expand in a two-dimensional (2D) culture system. Variations to this protocol, including three-dimensional (3D) culture systems, media supplements, plate coating to improve adhesion, and alternate disaggregation methods, could improve this technique and the overall success rate of establishing a tumor line. Overall, this protocol provides a base method for establishing and characterizing tumor cells from this rare tumor.
AB - Current methodologies for the expansion of primary tumor cell lines from rare tumor types are lacking. This protocol describes methods to expand primary tumor cells from surgically resected, malignant pleural mesothelioma (MPM) by providing a complete overview of the process from digestion to enrichment, expansion, cryopreservation, and phenotypic characterization. In addition, this protocol introduces concepts for tumor generation that may be useful for multiple tumor types such as differential trypsinization and the impact of dissociation methods on the detection of cell surface markers for phenotypic characterization. The major limitation of this study is the selection of tumor cells that will expand in a two-dimensional (2D) culture system. Variations to this protocol, including three-dimensional (3D) culture systems, media supplements, plate coating to improve adhesion, and alternate disaggregation methods, could improve this technique and the overall success rate of establishing a tumor line. Overall, this protocol provides a base method for establishing and characterizing tumor cells from this rare tumor.
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U2 - 10.3791/63374
DO - 10.3791/63374
M3 - Article
C2 - 35532258
AN - SCOPUS:85129503716
SN - 1940-087X
VL - 2022
JO - Journal of Visualized Experiments
JF - Journal of Visualized Experiments
IS - 182
M1 - e63374
ER -