3D Spheroids for Personalized Approach in Cancer Therapy

Introduction

Personalized approach of cancer therapy based on 3D spheroids offers a possibility of extensive analysis of the molecular characteristics of tumor cells and tumor sensitivity assays to various available drugs. One of the ways to emulate tumor tissue is 3D cell cultures in this case so-called 3D tumor spheroids. The effect of drugs observed on 3D cultures is often very different from the effects observed in 2D culture and is very close to the actual effect of the drug on the tumor in vivo. With these 3D structures we tested the function of dendrimers, which were tested on 2D culture too.

Material and method

Test of cytotoxicity was made. For the cell viability assay were used three different cell lines - B14, NRK and BRL. There were analyzed six dendrimers types in three generations (G1-G3) measured by the same methods – MTT and CV. Every cell line was exposed to concentration gradient. For culturing of 3D spheroids were used cell lines MCF-7 and CRL. The concentration of cells for forming spheroids was 547,000 per well. In three days is possible to observe spheroids. In the first step was made a test of toxicity on these spheroids with a nanostructure with drug delivering potential called dendrimers. These types of carbosilane nanoparticles were applied to spheroids in several concentrations. Spheroids were monitored for 4 days and their growth was analyzed by a measurement of their volume. The toxic effect of dendrimer was evaluated using growth curves and analysis of ATP production by the luminescence method using CellTiter-Glo® Luminescent Cell Viability Assay.

Results and discussion

Cytotoxicity of PMe₃ was similar or slightly lower as compared to NMe₃ dendrimers. The highest toxicity of Pbu₃ on 2D model was detected. Dendrimers with lowest cytotoxicity were P(Et₂)₂(CH₂)₃OH and P(Ph)₃ . Also was observed their accumulation in mitochondria. Their low toxicity was confirmed by tests on 3D spheroids too. Therefore, both dendrimers could represent unique drug deliver systems for mitochondria targeting.

Conclusions

Toxicity of these two dendrimers P(Ph)₃ and P(Et₂)₂(CH₂)₃OH are already tested on the 2D cultures and 3D spheroids. Results of both models will be compared.

Research was supported by project 2018-53-005-3 Internal Grand Agency UJEP.