Three dimensional models of dedifferentiated liposarcoma cell lines: scaffold-based and scaffold-free approaches

by Selleckchem | Jun 20 2023

Sarcomas are rare malignancies, the number of reports is limited, and this rarity makes further research difficult even though liposarcoma is one of major sarcomas. 2D cell culture remains an important role in establishing basic tumor biology research, but its various shortcomings and limitations are still of concern, and it is now well-accepted that the behavior of 3D-cultured cells is more reflective of in vivo cellular responses compared to 2D models. This study aimed to establish 3D cell culture of liposarcomas using two different methods: scaffold-based (Matrigel extracellular matrix [ECM] scaffold method) and scaffold-free (Ultra-low attachment [ULA] plate). Lipo246, Lipo224 and Lipo863 cell lines were cultured, and distinctive differences in structures were observed in Matrigel 3D model: Lipo224 and Lipo863 formed spheroids, whereas Lipo246 grew radially without forming spheres. In ULA plate approaches, all cell lines formed spheroids, but Lipo224 and Lipo863 spheroids showed bigger size and looser aggregation than Lipo246. Formalin fixed, paraffin embedded (FFPE) blocks were obtained from all 3D models, confirming the spheroid structures. The expression of MDM2, Ki-67 positivity and MDM2 amplification were confirmed by IHC and DNAscope™, respectively. Protein and DNA were extracted from all samples and MDM2 upregulation was confirmed by western blot and qPCR analysis. After treatment with MDM2 inhibitor SAR405838, DDLPS spheroids demonstrated different sensitivity patterns from 2D models. Taken together, we believed that 3D models would have a possibility to provide us a new predictability of efficacy and toxicity, and considered as one important process in in vitro pre-clinical phase prior to moving forward to clinical trials.

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The passage describes a study that aimed to establish 3D cell culture models of liposarcomas, a type of rare malignant tumor. The researchers used two different methods: a scaffold-based approach using Matrigel extracellular matrix (ECM) and a scaffold-free approach using an Ultra-low attachment (ULA) plate.

The study used three different liposarcoma cell lines: Lipo246, Lipo224, and Lipo863. In the Matrigel 3D model, Lipo224 and Lipo863 formed spheroids, while Lipo246 grew radially without forming spheres. In the ULA plate approach, all three cell lines formed spheroids, but Lipo224 and Lipo863 spheroids were larger in size and had looser aggregation compared to Lipo246.

Formalin-fixed, paraffin-embedded (FFPE) blocks were obtained from all 3D models, confirming the presence of spheroid structures. The expression of MDM2, a protein of interest, was examined using immunohistochemistry (IHC), and MDM2 amplification was analyzed using DNAscope™. Protein and DNA were extracted from the samples, and MDM2 upregulation was confirmed through western blot and qPCR analysis.

After treating the liposarcoma spheroids with an MDM2 inhibitor called SAR405838, the researchers observed different sensitivity patterns compared to the 2D cell culture models.

The study concludes that 3D cell culture models have the potential to provide new insights into the efficacy and toxicity of treatments. These models can be considered an important step in the in vitro pre-clinical phase before moving on to clinical trials. By using 3D models, researchers hope to improve the understanding of liposarcoma biology and develop more effective treatment strategies for this rare malignancy.