Transient transfection induces different intracellular calcium signaling in CHO K1 versus HEK 293 cells.
For the controlled production of recombinant proteinsin mammalian cells by transient transfection, it maybe desirable not only to manipulate, but also todiagnose the expression success early. Here, weapplied laser scanning confocal microscopy to monitortransfection induced intracellular Ca(2+)responses. We compared Chinese hamster ovary (CHO K1)versus human embryo kidney (HEK) 293 cell lines, whichdiffer largely in their transfectability. An improvedcalcium phosphate transfection method was used for itssimplicity and its demonstrated upscale potential.Cytosolic Ca(2+) signaling appeared to inverselyreflect the cellular transfection fate. Virtually allCHO cells exhibited asynchronous, cytosolicCa(2+) oscillations, which peaked 4 h afteraddition of the transfecting solution. Yet, most ofthe HEK cells displayed a slow and continuousCa(2+) increase over the time of transfection. CHOcells, when exposed to a transfection-enhancingglycerol shock, strongly downregulated their Ca(2+)response, including its oscillations. When treatedwith thapsigargin, a Ca(2+) store depleting drug,the number of successfully transfected CHO cells was significantly reduced. Our result points tointracellular store release as a critical componentfor the transfection fate of CHO cells, and its early detection before product visualization.
Preuss
A K
AK
Laboratory of Physical Chemistry of Polymers and Membranes, Chemistry Department, Swiss Federal Institute of Technology, CH 1015, Lausanne, Switzerland.
Connor
J A
JA
Vogel
H
H