BACKGROUND: Pulsed electric field has been widely used to facilitate molecular cargo transfer into cells. However, the cell viability is often decreased when trying to increase the electrotransfer efficiency. We hypothesize that the decrease is due to electropermeabilization of cell membrane that disrupts homeostasis of intracellular microenvironment. Thus, a reduction in the membrane permeabilization may increase the cell viability. MATERIALS AND METHODS: Different compounds were supplemented into the pulsing buffer prior to electrotransfer for reduction of cell membrane damage. Extent of the damage was quantified by leakiness of the membrane to a fluorescent dye, calcein, preloaded into cells. At 24 hours post electrotransfer, cell viability and electrotransfer efficiency were quantified with flow cytometry. RESULTS: The cell viability could be substantially increased by supplementation of either type B gelatin or bovine serum albumin (BSA), without compromising the electrotransfer efficiency. The supplementation also decreased the amount of calcein leaking out of the cells, suggesting that the improvement in cell viability was due to the reduction in electrotransfer-induced membrane damage. CONCLUSION: Data from the study demonstrate that type B gelatin and BSA can be used as inexpensive supplements for improving cell viability in electrotransfer.
BACKGROUND: Pulsed electric field has been widely used to facilitate molecular cargo transfer into cells. However, the cell viability is often decreased when trying to increase the electrotransfer efficiency. We hypothesize that the decrease is due to electropermeabilization of cell membrane that disrupts homeostasis of intracellular microenvironment. Thus, a reduction in the membrane permeabilization may increase the cell viability. MATERIALS AND METHODS: Different compounds were supplemented into the pulsing buffer prior to electrotransfer for reduction of cell membrane damage. Extent of the damage was quantified by leakiness of the membrane to a fluorescent dye, calcein, preloaded into cells. At 24 hours post electrotransfer, cell viability and electrotransfer efficiency were quantified with flow cytometry. RESULTS: The cell viability could be substantially increased by supplementation of either type B gelatin or bovine serum albumin (BSA), without compromising the electrotransfer efficiency. The supplementation also decreased the amount of calcein leaking out of the cells, suggesting that the improvement in cell viability was due to the reduction in electrotransfer-induced membrane damage. CONCLUSION: Data from the study demonstrate that type B gelatin and BSA can be used as inexpensive supplements for improving cell viability in electrotransfer.
Authors: Theodore L Roth; Cristina Puig-Saus; Ruby Yu; Eric Shifrut; Julia Carnevale; P Jonathan Li; Joseph Hiatt; Justin Saco; Paige Krystofinski; Han Li; Victoria Tobin; David N Nguyen; Michael R Lee; Amy L Putnam; Andrea L Ferris; Jeff W Chen; Jean-Nicolas Schickel; Laurence Pellerin; David Carmody; Gorka Alkorta-Aranburu; Daniela Del Gaudio; Hiroyuki Matsumoto; Montse Morell; Ying Mao; Min Cho; Rolen M Quadros; Channabasavaiah B Gurumurthy; Baz Smith; Michael Haugwitz; Stephen H Hughes; Jonathan S Weissman; Kathrin Schumann; Jonathan H Esensten; Andrew P May; Alan Ashworth; Gary M Kupfer; Siri Atma W Greeley; Rosa Bacchetta; Eric Meffre; Maria Grazia Roncarolo; Neil Romberg; Kevan C Herold; Antoni Ribas; Manuel D Leonetti; Alexander Marson Journal: Nature Date: 2018-07-11 Impact factor: 49.962