Professor Rutgers University New York, New York, United States
Introduction:: Introduction: The retina is a crucial component of the visual system that contains photoreceptor cells responsible for transmitting visual information to the brain. However, the loss of these photoreceptor cells is a common outcome of several eye conditions, such as age-related macular degeneration, diabetic retinopathy, and retinitis pigmentosa, leading to a need for replacement cells for up to 197.5 million people globally. Transplantation of retinal progenitor cells (RPCs) has produced exciting potential to restore vision via cell replacement therapy. Our lab has previously developed microfluidic systems able to induce migration via applied chemical gradient. In this project, we demonstrate that pulsed electric fields in combination with chemical gradients increase the directionality and distance of RPC migration. Experiments illustrate possible therapeutic benefits of translating these electro-chemical fields in retinal explants and eventually in vivo.
Materials and Methods:: Materials and Methods: Rat retinal progenitor cells (R28) were used for this model. The microfluidic device was coated in laminin for 24 hours for cells to be seeded, then another 24 hours was passed before establishing the electrochemical gradient in order for cells to attach. The microfluidic device was made by curing PDMS in a mold. The device features two reservoirs connected by a microchannel of 200μm in diameter, where cells are able to be identified with a microscope to determine migration velocity. Chemical gradients are able to be established by controlling the concentration of chemoattractants in both reservoirs. The device has two stainless steel needles installed which serve as electrodes. R28 migration velocity was measured in the device over time.
Results, Conclusions, and Discussions:: Results: Migration velocity was calculated from multiple images over time. The control condition had no significant MV, and the electrical, chemical, and combination conditions all had significant MV. Comparisons show that the combination field condition significantly had the greatest MV.
Conclusion: In this work we developed a reliable in vitro method to assess influences of electrochemical fields cells on transplantable cells. The study ultimately aims to provide novel insight on R28 migration and the possible use of electrochemical fields for clinical settings to develop targeted therapies that aim to advance photoreceptor cell replacement therapies.
