Increased transduction efficiency in microfluidic devices
Increased transduction efficiency in microfluidic devices
Arezoo Karamivandishi,1Masoud Soleimani,2,*
1. Tarbiat modares medical university 2. Tarbiat modares medical university. & shahid Beheshti medical university
Introduction: Gene therapy using lentivectors (LV) is a promising approach to treat many disorders such as multiple myeloma (MM). Multiple myeloma is a second most common malignancy that characterized by the accumulation of malignant plasma cells in the bone marrow. In this study, we targeted special antigen in MM, B- cell maturation antigen by using the droplet-based microfluidic device. There are limitations in manufacturing LV due to inefficient cell transduction. To overcome this problem, we designed a focused-flow droplet generator device to increase gene transfer efficiency.
Methods: Device designed by soft lithography technique. Human myeloma cells were transduced by LVV in our designed chip and plates as control. Transduction efficiency is assessed by determining GFP expression on FACS.
Results: Transduction efficiency was assessed at decreasing virus concentration (MOI) while keeping picodroplet size constant. Transduction efficiency in picodroplets was higher than controls in human myeloma cells at all MOI s tested.
Conclusion: transduction efficiencies in pico droplets are comparable to levels obtained with transduction in conventional methods, indicating efficient gene delivery to target cells. Overall, our study provides a novel approach to the field of gene transfer to cells, especially transfection -resistant cells.