 |
 |
 |
 |
||
|
 |
|||||||||||||||||||||||
 | ||||||||||||||||||||||||
 | ||||||||||||||||||||||||
 | ||||||||||||||||||||||||
 | ||||||||||||||||||||||||
Confocal Microscopy Image Gallery
Normal African Green Monkey Kidney Fibroblast Cells (CV-1 Line)
Investigators working with simian virus 40 (SV40) or in the field of AIDS research often utilize CV-1 cells, which were initially used in studies of the Rous sarcoma virus (RSV). A team led by F. C. Jensen established the CV-1 cell line in the early 1960s.
The cells from which the line was initiated were sampled from the renal tissue of an African green monkey (Cercopithecus aethiops). CV-1 cells are popularly employed in transfection experiments and are known to be susceptible to a wide array of viruses, including several types of encephalitis (California, Eastern, and Western), poliovirus 1, and herpes simplex, as well as SV40. The fibroblasts grow rapidly in culture. At high passage levels, shifts in the number of chromosomes the cells contain may occur.
Kidney cell lines initiated from African green monkeys and other non-human primates have played an important role in vaccine production. Their use for this purpose, however, has been the subject of a considerable amount of controversy since it is possible for humans to be exposed to simian viruses through such means. Stringent testing for viral contamination is carried out on all cell lines used to generate vaccines today, but some individuals are still concerned about the practice. Underlying this concern is the fact that in the 1960s it was discovered that millions of people were exposed to the polyoma virus SV40 through polio vaccinations. SV40 can cause cancer in laboratory animals, but its effect on humans is not known with certainty. A number of researchers are currently studying tumor samples in order to determine if there is any link between the virus and human cancers.
The adherent culture of CV-1 kidney fibroblasts featured above was immunofluorescently labeled with anti-vinculin mouse monoclonal primary antibodies followed by goat anti-mouse IgG secondary antibodies conjugated to Alexa Fluor 647 (pseudocolored blue). In addition, the specimen was stained for DNA with the ultraviolet-absorbing probe Hoechst 33342 (pseudocolored cyan), for the cytoskeletal filamentous actin network with Alexa Fluor 488 conjugated to phalloidin, and for mitochondria with MitoTracker Red CMXRos. Images were recorded with a 60x oil immersion objective using a zoom factor of 1.5 and sequential scanning with the 405-nanometer spectral line of a blue diode laser, 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.
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.