Protocol for generation of 3D bone marrow surrogate microenvironments in a rotary cell culture system

In this protocol, we describe how to generate 3D culture surrogates of chronic lymphocytic leukemia (CLL) and multiple myeloma (MM) bone marrow microenvironments. We detail the use of culturing scaffolds populated with BM stromal cells and tumor cells in the RCCS™ bioreactor. This 3D culture can eff...

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Bibliographic Details
Published in:STAR protocols 2022-09, Vol.3 (3), p.101601-101601, Article 101601
Main Authors: Belloni, Daniela, Ferrarini, Marina, Ferrero, Elisabetta, Guzzeloni, Virginia, Barbaglio, Federica, Ghia, Paolo, Scielzo, Cristina
Format: Article
Language:English
Subjects:
Biotechnology and bioengineering
Cancer
Cell Biology
Cell culture
Immunology
Protocol
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Summary:In this protocol, we describe how to generate 3D culture surrogates of chronic lymphocytic leukemia (CLL) and multiple myeloma (MM) bone marrow microenvironments. We detail the use of culturing scaffolds populated with BM stromal cells and tumor cells in the RCCS™ bioreactor. This 3D culture can efficiently recapitulate tumor-stroma crosstalk and allows the testing of drugs such as ibrutinib and bortezomib. Moreover, this protocol can be used for the generation of other and more complex tumor microenvironments. For complete details on the use and execution of this protocol, please refer to Belloni et al. (2018) and Barbaglio et al. (2021). [Display omitted] •Step-by-step generation of a 3D bone marrow microenvironment for CLL and MM•Set up of a dynamic 3D culture in an RCCS™ bioreactor•Integrative analysis of the 3D scaffolds in response to drug treatment Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. In this protocol, we describe how to generate 3D culture surrogates of chronic lymphocytic leukemia (CLL) and multiple myeloma (MM) bone marrow microenvironments. We detail the use of culturing scaffolds populated with BM stromal cells and tumor cells in the RCCS™ bioreactor. This 3D culture can efficiently recapitulate tumor-stroma crosstalk and allows the testing of drugs such as ibrutinib and bortezomib. Moreover, this protocol can be used for the generation of other and more complex tumor microenvironments.
ISSN:2666-1667
2666-1667
DOI:10.1016/j.xpro.2022.101601