Methods for Establishing a Renal Cell Carcinoma Tumor Spheroid Model With Immune Infiltration for Immunotherapeutic Studies

• DOI: 10.3389/fonc.2022.898732
• Publication Date: 2022-07-28T06:13:53Z
• Authors: Leonard Lugand; Guillaume Mestrallet; Rebecca Laboureur; Clement Dumont; Fatiha Bouhidel; Malika Djouadou; Alexandra Masson-...; Francois Desgrand...; Stephane Culine; Edgardo D. Carosella; Nathalie Rouas-Fr...; Joel LeMaoult
• Keywords: Spheroid

Tumor spheroids play an increasingly important role in cancer research. Their ability to recapitulate crucial features of tumor biology that are lost the classically used 2D models along with their relative simplicity and handiness have made them most studied 3D model. application as a theranostic tool or means study tumor-host interaction is now well-established various cancers. However, use field Renal Cell Carcinoma (RCC) remains very limited. The aim this work present methods implement basic RCC spheroid These cover steps from dissociation infiltration by immune cells. We protocol for using Liberase TM introduce culture medium containing Epithelial Growth Factor Hydrocortisone allowing faster growth primary show liquid overlay technique allows formation cell lines cultures. method morphological criteria select homogeneous population based on Fiji macro. then can be infiltrated PBMCs after activation OKT3 IL-15. Finally, we provide example implementing killing assay observing increased destruction treatment PD-1 inhibitors. Thus straightforward presented here allow efficient simple model standardized infused cells immunotherapies.