[Using GFP retrovirus to label tumor cells and vascular endothelia cells].
BACKGROUND
To prepare retrovirus which carry GFP gene and are able to label living cells simply and rapidly.


METHODS
The recombinant retroviral vector pLNCX-GFP was constructed by inserting 780?bp GFP cDNA fragment into the MCS site of retroviral plasmid pLNCX. Both ecotropic packaging cell line ΦX-Eco and amphotropic packaging cell line ΦX-Ampho and PA317 were transfected by pLNCX-GFP with liposome. The supernate collected from transfected packaging cells was used to infect a variety of tumor cell lines and vascular endothelia cell lines.


RESULTS
When packaging cells were transfected by retroviral vector pLNCX-GFP, the GFP expression could be observed in 25%-40% of cells and GFP retrovirus then could be detected, however G418 resistant clones showed more stable GFP expression and higher retrovirus titer. The GFP retrovirus from different packaging cell line showed variant ability to infect tumor cell lines and vascular endothelia cell lines and the tumor cells infected by GFP retrovirus showed stable GFP expression in vitro. GFP transduced tumor cells could grow in syngenic animal and continue expressing GFP.


CONCLUSIONS
Using GFP retrovirus to label target cells represent an important advantage over conventional plasmid because they can efficiently transfer GFP gene into target cells and GFP can be stably expressed in target cells no matter in vitro or in vivo.
Central Laboratory, The First People's Hospital of Shanghai, Shanghai 200080, P.R.China.
K
K
Xie
P
P
Xu
Q
Q
Gu
W
W
Liu
F
F
Wang
Y
Y
Tian
X
X
Chen
C
C
Li
Q
Q
Huang