Exosome isolation with microfluidic system and development of drug carrier system targeting cancer cells

Objective: In breast cancer, triple negative breast cancer is an aggressive subtype with a poor clinical prognosis. Exosomes are small-sized vesicles produced naturally by cells for communication purposes, and they also attract attention as a drug carrier system. This study aimed to investigate the isolation of exosomes secreted from Jurkat T cells with a microfluidic (MF) system and the effects of paclitaxel (PTX) loading on exosomes on breast cancer. Materials and methods: Exosomes were isolated from Jurkat T-cells by stimulating exosome relase using MF and ultracentrifugation. After size analysis and characterization (CD63 and CD81) of exosomes were performed, PTX was loaded by electroporation. Cell viability of Exo-PTX on MDA-MB-231 cells was analyzed by MTT method, and its apoptotic effect was analyzed by flow cytometry with Annexin V-FITC-PI and JC-1 staining. The effect of Exo-PTX on migration ability was determined by the Boyden Chamber method and its role on chemokines was determined by CXCR4/CXCL12 gene expression analysis. Results: Exosomes isolated from Jurkat T cells were determined to be below 100 nm in size and carry CD63 and CD81 markers. Exo-PTX (10μg/mL) and PTX (15 nM) and above concentrations significantly reduced cell viability, there was a significant increase in early and late apoptosis, decreased migration abilities and suppressed the gene expression levels of CXCR4/CXCL12 determined. Conclusion: Isolation of exosomes of fixed size in MF systems from suspended cells such as T lymphocytes can offers an advantage. Jurkat exosomes can be used in the treatment of metastatic cancers to maintain the effectiveness of chemotherapeutic agents and ensure their entry into the cell. Keywords: Exosome, Jurkat T cell, Breast Cancer, Microfluidic system, Paclitaxel