Olympus Olympus BX2 Research Microscopes .
Laser Scanning Confocal Microscopy
Confocal Microscopy Image Gallery

Rhesus Monkey Kidney Epithelial Cells (LLC-MK2 Line)

LLC-MK2 is an epithelial line that was established in the 1950s from a pooled suspension prepared from renal tissue excised from six rhesus monkeys (Macaca mulatta). LLC-MK2 cellular products include the protease plasminogen activator associated with the kidneys that normally instigates the process of fibrinolysis by converting plasminogen to plasmin.

LLC-MK2 Cells

The LLC-MK2 line demonstrates susceptibility to polioviruses 1, 2, and 3 and tests negative for reverse transcriptase, indicating the lack of integral retrovirus genomes. The cells are a common host used in transfection experiments. In culture, LLC-MK2 cells grow adherently to both glass and polymer surfaces.

LLC-MK2 cells have been utilized in mumps vaccine production and in the isolation of parainfluenza viruses. Rhesus monkeys, from which the LLC-MK2 line was derived, have contributed to science in a number of other ways as well. The Old World primates, which have a close genetic relationship to humans and are relatively easy to rear in captivity, have been used as subjects for laboratory research for many years. In the 1940s, work with rhesus monkeys resulted in the discovery of the Rh factor, and the following decade they were employed in famous psychological experiments on maternal deprivation. More recently, rhesus monkeys have been utilized in groundbreaking cloning and transgenic experiments.

The rhesus monkey kidney cells presented in this confocal image were resident in an adherent culture stained for F-actin with BODIPY FL conjugated to phallacidin, and for mitochondria with MitoTracker Red CMXRos (pseudocolored yellow). In addition, the specimen was counterstained for DNA with the red-absorbing dye TO-PRO-3. Images were recorded with a 60x oil immersion objective using a zoom factor of 2.5 and sequential scanning with the 488-nanometer spectral line of an argon-ion laser, the 543-nanometer line from a green helium-neon laser, and the 633-nanometer line of a red helium-neon laser. During the processing stage, individual image channels were pseudocolored with RGB values corresponding to each of the fluorophore emission spectral profiles unless otherwise noted above.

Additional Confocal Images of Rhesus Monkey Kidney Epithelial (LLC-MK2) Cells

Distribution of F-Actin and Mitochondria in LLC-MK2 Cells - A triple fluorophore combination of MitoTracker Red CMXRos, BODIPY FL conjugated to phallacidin, and TO-PRO-3 was used to label an adherent log phase culture of LLC-MK2 cells for mitochondria, the filamentous actin network, and nuclear DNA, respectively. The cells were first treated with the MitoTracker probe in growth medium for one hour, washed and fixed with paraformaldehyde (prepared in growth medium), permeabilized, and blocked with bovine serum albumen. The cells were subsequently labeled with the conjugated phallacidin and counterstained with the cyanine monomer (TO-PRO-3) reagent.

Contributing Authors

Nathan S. Claxton, Shannon H. Neaves, and Michael W. Davidson - National High Magnetic Field Laboratory, 1800 East Paul Dirac Dr., The Florida State University, Tallahassee, Florida, 32310.