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Application Note: Cationic Polymer Transfections In Corning® HYPERFlask® Cell Culture Vessels
By Pilar Pardo, Todd Upton, Ph.D. Corning Incorporated, Life Sciences
One of the most useful tools in cell biology research is transfection, the introduction of foreign DNA into eukaryotic cells. Transient transfection of mammalian cell lines is a popular technique used to express genes of interest for a brief period of time, typically 2 to 3 days. In today's industry, there is an increasingly growing need to carry out these transfections in larger quantities of cells.
The Corning HYPERFlask Cell Culture Vessel, a high yield, high performance flask that utilizes a multilayered gas permeable growing surface for efficient gas exchange, can help researchers transfect larger amounts of cells. In this study, HeLa and CHO-K1 cells were transfected using Polyplus-transfection jetPEI™ transfection reagent along with two different DNA constructs: one designed to test transfection via secreted protein production and the other to determine transfection efficiency through GFP expression within cells.
jetPEI™, a polyethylenimine (PEI) derivative, is a proprietary transfection reagent that binds to DNA through electrostatic forces for a cationic polymer transfection. The jetPEI/DNA complex gently enters the cell by endocytosis, where the DNA is then released into the cytoplasm. The jetPEI reagent is water-soluble and can be used in the presence of serum in culture media, eliminating the need for medium changes before or after transfection, making this method ideally suited for large scale transfections when using the Corning® HYPERFlask® vessel.
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