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Methods for Establishing a Renal Cell Carcinoma Tumor Spheroid Model With Immune Infiltration for Immunotherapeutic Studies
Tumor spheroids play an increasingly important role in cancer research. Their ability to recapitulate crucial features of tumor biology that are lost in the classically used 2D models along with their relative simplicity and handiness have made them the most studied 3D tumor model. Their application...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9366089/ https://www.ncbi.nlm.nih.gov/pubmed/35965544 http://dx.doi.org/10.3389/fonc.2022.898732 |
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author | Lugand, Leonard Mestrallet, Guillaume Laboureur, Rebecca Dumont, Clement Bouhidel, Fatiha Djouadou, Malika Masson-Lecomte, Alexandra Desgrandchamps, Francois Culine, Stephane Carosella, Edgardo D. Rouas-Freiss, Nathalie LeMaoult, Joel |
author_facet | Lugand, Leonard Mestrallet, Guillaume Laboureur, Rebecca Dumont, Clement Bouhidel, Fatiha Djouadou, Malika Masson-Lecomte, Alexandra Desgrandchamps, Francois Culine, Stephane Carosella, Edgardo D. Rouas-Freiss, Nathalie LeMaoult, Joel |
author_sort | Lugand, Leonard |
collection | PubMed |
description | Tumor spheroids play an increasingly important role in cancer research. Their ability to recapitulate crucial features of tumor biology that are lost in the classically used 2D models along with their relative simplicity and handiness have made them the most studied 3D tumor model. Their application as a theranostic tool or as a means to study tumor-host interaction is now well-established in various cancers. However, their use in the field of Renal Cell Carcinoma (RCC) remains very limited. The aim of this work is to present methods to implement a basic RCC spheroid model. These methods cover the steps from RCC tumor dissociation to spheroid infiltration by immune cells. We present a protocol for RCC dissociation using Liberase TM and introduce a culture medium containing Epithelial Growth Factor and Hydrocortisone allowing for faster growth of RCC primary cells. We show that the liquid overlay technique allows for the formation of spheroids from cell lines and from primary cultures. We present a method using morphological criteria to select a homogeneous spheroid population based on a Fiji macro. We then show that spheroids can be infiltrated by PBMCs after activation with OKT3 or IL-15. Finally, we provide an example of application by implementing an immune spheroid killing assay allowing observing increased spheroid destruction after treatment with PD-1 inhibitors. Thus the straightforward methods presented here allow for efficient spheroid formation for a simple RCC 3D model that can be standardized and infused with immune cells to study immunotherapies. |
format | Online Article Text |
id | pubmed-9366089 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-93660892022-08-12 Methods for Establishing a Renal Cell Carcinoma Tumor Spheroid Model With Immune Infiltration for Immunotherapeutic Studies Lugand, Leonard Mestrallet, Guillaume Laboureur, Rebecca Dumont, Clement Bouhidel, Fatiha Djouadou, Malika Masson-Lecomte, Alexandra Desgrandchamps, Francois Culine, Stephane Carosella, Edgardo D. Rouas-Freiss, Nathalie LeMaoult, Joel Front Oncol Oncology Tumor spheroids play an increasingly important role in cancer research. Their ability to recapitulate crucial features of tumor biology that are lost in the classically used 2D models along with their relative simplicity and handiness have made them the most studied 3D tumor model. Their application as a theranostic tool or as a means to study tumor-host interaction is now well-established in various cancers. However, their use in the field of Renal Cell Carcinoma (RCC) remains very limited. The aim of this work is to present methods to implement a basic RCC spheroid model. These methods cover the steps from RCC tumor dissociation to spheroid infiltration by immune cells. We present a protocol for RCC dissociation using Liberase TM and introduce a culture medium containing Epithelial Growth Factor and Hydrocortisone allowing for faster growth of RCC primary cells. We show that the liquid overlay technique allows for the formation of spheroids from cell lines and from primary cultures. We present a method using morphological criteria to select a homogeneous spheroid population based on a Fiji macro. We then show that spheroids can be infiltrated by PBMCs after activation with OKT3 or IL-15. Finally, we provide an example of application by implementing an immune spheroid killing assay allowing observing increased spheroid destruction after treatment with PD-1 inhibitors. Thus the straightforward methods presented here allow for efficient spheroid formation for a simple RCC 3D model that can be standardized and infused with immune cells to study immunotherapies. Frontiers Media S.A. 2022-07-28 /pmc/articles/PMC9366089/ /pubmed/35965544 http://dx.doi.org/10.3389/fonc.2022.898732 Text en Copyright © 2022 Lugand, Mestrallet, Laboureur, Dumont, Bouhidel, Djouadou, Masson-Lecomte, Desgrandchamps, Culine, Carosella, Rouas-Freiss and LeMaoult https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Oncology Lugand, Leonard Mestrallet, Guillaume Laboureur, Rebecca Dumont, Clement Bouhidel, Fatiha Djouadou, Malika Masson-Lecomte, Alexandra Desgrandchamps, Francois Culine, Stephane Carosella, Edgardo D. Rouas-Freiss, Nathalie LeMaoult, Joel Methods for Establishing a Renal Cell Carcinoma Tumor Spheroid Model With Immune Infiltration for Immunotherapeutic Studies |
title | Methods for Establishing a Renal Cell Carcinoma Tumor Spheroid Model With Immune Infiltration for Immunotherapeutic Studies |
title_full | Methods for Establishing a Renal Cell Carcinoma Tumor Spheroid Model With Immune Infiltration for Immunotherapeutic Studies |
title_fullStr | Methods for Establishing a Renal Cell Carcinoma Tumor Spheroid Model With Immune Infiltration for Immunotherapeutic Studies |
title_full_unstemmed | Methods for Establishing a Renal Cell Carcinoma Tumor Spheroid Model With Immune Infiltration for Immunotherapeutic Studies |
title_short | Methods for Establishing a Renal Cell Carcinoma Tumor Spheroid Model With Immune Infiltration for Immunotherapeutic Studies |
title_sort | methods for establishing a renal cell carcinoma tumor spheroid model with immune infiltration for immunotherapeutic studies |
topic | Oncology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9366089/ https://www.ncbi.nlm.nih.gov/pubmed/35965544 http://dx.doi.org/10.3389/fonc.2022.898732 |
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