Technical report: liquid overlay technique allows the generation of homogeneous osteosarcoma, glioblastoma, lung and prostate adenocarcinoma spheroids that can be used for drug cytotoxicity measurements

Frontiers in Bioengineering and Biotechnology

Bioengineering and Biotechnology

Denis Cochonneau¹  Emilie Ollivier¹  François Vallette²  Javier Muñoz-Garcia³  Marie-Françoise Heymann³  Camille Jubelin⁴  Dominique Heymann⁵  Lisa Oliver⁶ 

[1] Institut de Cancérologie de l’Ouest, Tumor Heterogeneity and Precision Medicine Lab, Saint-Herblain, France;Institut de Cancérologie de l’Ouest, Tumor Heterogeneity and Precision Medicine Lab, Saint-Herblain, France;Nantes Université, Institut National de la Santé et de la Recherche Médicale (INSERM), Centre de Recherche en Cancérologie et Immunologie Nantes-Angers (CRCINA), Nantes, France;Nantes Université, Centre National de la Recherche Scientifique (CNRS), Unit in Biological Sciences and Biotechnologies (US2B), Nantes, France;Institut de Cancérologie de l’Ouest, Tumor Heterogeneity and Precision Medicine Lab, Saint-Herblain, France;Nantes Université, Centre National de la Recherche Scientifique (CNRS), Unit in Biological Sciences and Biotechnologies (US2B), Nantes, France;Institut de Cancérologie de l’Ouest, Tumor Heterogeneity and Precision Medicine Lab, Saint-Herblain, France;Atlantic Bone Screen, Saint-Herblain, France;Nantes Université, Centre National de la Recherche Scientifique (CNRS), Unit in Biological Sciences and Biotechnologies (US2B), Nantes, France;Institut de Cancérologie de l’Ouest, Tumor Heterogeneity and Precision Medicine Lab, Saint-Herblain, France;Department of Oncology and Metabolism, Medical School, University of Sheffield, Sheffield, United Kingdom;Nantes Université, Institut National de la Santé et de la Recherche Médicale (INSERM), Centre de Recherche en Cancérologie et Immunologie Nantes-Angers (CRCINA), Nantes, France;CHU de Nantes, Nantes, France;

关键词: 3D culture (three-dimensional spheroids);    homogeneous spheroid model;    liquid overlay technique;    hanging drop;    lung adenocarcinoma;    prostate adenocarcinoma;    osteosarcoma;    glioblastoma;   

DOI  :  10.3389/fbioe.2023.1260049

 received in 2023-07-17, accepted in 2023-09-25,  发布年份 2023

来源: Frontiers

【 摘 要 】

Introduction: The mechanisms involved in cancer initiation, progression, drug resistance, and disease recurrence are traditionally investigated through in vitro adherent monolayer (2D) cell models. However, solid malignant tumor growth is characterized by progression in three dimensions (3D), and an increasing amount of evidence suggests that 3D culture models, such as spheroids, are suitable for mimicking cancer development. The aim of this report was to reaffirm the relevance of simpler 3D culture methods to produce highly reproducible spheroids, especially in the context of drug cytotoxicity measurements.Methods: Human A549 lung adenocarcinoma, LnCaP prostate adenocarcinoma, MNNG/HOS osteosarcoma and U251 glioblastoma cell lines were grown into spheroids for 20 days using either Liquid Overlay Technique (LOT) or Hanging Drop (HD) in various culture plates. Their morphology was examined by microscopy. Sensitivity to doxorubicin was compared between MNNG/HOS cells grown in 2D and 3D.Results: For all cell lines studied, the morphology of spheroids generated in round-bottom multiwell plates was more repeatable than that of those generated in flat-bottom multiwell plates. HD had no significant advantage over LOT when the spheroids were cultured in round-bottom plates. Finally, the IC50 of doxorubicin on MNNG/HOS cultured in 3D was 18.8 times higher than in 2D cultures (3D IC50 = 15.07 ± 0.3 µM; 2D IC50 = 0.8 ± 0.4 µM; *p < 0.05).Discussion: In conclusion, we propose that the LOT method, despite and because of its simplicity, is a relevant 3D model for drug response measurements that could be scaled up for high throughput screening.

【 授权许可】

Unknown   
Copyright © 2023 Jubelin, Muñoz-Garcia, Cochonneau, Ollivier, Vallette, Heymann, Oliver and Heymann.