Arch Virol (2002) 147: 321–333
Generation of a life-expanded rhesus monkey ﬁbroblast
cell line for the growth of rhesus rhadinovirus (RRV)
, S. Wong
, S. St. Jeor
, and G. S. Pari
Department of Microbiology,University of Nevada-Reno,Reno,Nevada,U.S.A.
Division of Pathobiology and Immunology,Oregon Health Sciences University/
Oregon Regional Primate Research Center,Beaverton,Oregon,U.S.A.
Accepted October 2,2001
Summary. RRV,the rhesus macaque equivalent to HHV-8 or kaposi’s sarcoma-
associated herpesvirus (KSHV) was recently isolated from a simian immuno-
deﬁciency virus (SIV) infected macaque with a lymphoproliferative disorder. The
growth of RRV in tissue culture requires propagation of primary rhesus monkey
ﬁbroblasts (RFs). In an effort to extend the life of these primary cells in tissue
culture,the catalytic subunit of telomerase (hTERT) was introduced into RF cells
using a recombinant retrovirus. This new cell line,Telo-RFs,have currently been
passed in tissue culture over 80 times compared to a maximum passage number
of 38 for wild type RFs,remain fully permissive for RRV DNA replication and
production of infectious virus.Viral gene expression of immediate-early and early
RNA transcripts was virtually identical to that observed in wild-type (wt) RFs.
In addition,transfection experiments show that telo-RFs are easily and more
efﬁciently transfected than wtRFs.
The study of many viruses requires the growth and maintenance of primary cells.
Some viruses,such as some of those in the herpesvirus family,can only be grown
in normal diploid ﬁbroblasts. In order to carry out experiments involving viral
attachment,permissive gene expression and viral DNA replication,the steady
supply of tissue to generate primary cell lines is needed. In many cases,acquiring
a supply of human or non-human primate tissue can be difﬁcult since viable
tissue can be limited. In addition,even when tissue is acquired and cell lines
are made,primary cell lines have as a major disadvantage a limited life in tissue
culture. Our goal was to increase the life span and availability of Rhesus Macaque
ﬁbroblasts to enhance the study of Rhesus Macaque Rhadinovirus (RRV) lytic