HEPATOCELLULAR TRANSPLANTATION AND TARGETING GENETIC MARKERS TO HEPATIC CELLS 
1 
I 
A. SOUTHERN BLOT PROBED WITH NEO-R B. NORTHERN BLOT PROBED WITH NEO-R 
j Figure 2, Expression of NEO-R in human hepatocvtes transduced with N2 or NASPAH (Peng 
et al. 1988) bv exposure to viral supernatant or co-cultivation. Southern and northern 
j blots of hviman or murine hepatocytes transduced with vector (N2 or NASPAH) probed with 
I NEO-R gene. This experiment compared transduction with vector containing supernantant 
i and cocultivation with vector producing cells followed by selection against the vector 
J producing cells in tyrosine free, hormonally defined media for 2 weeks (Ledley, 
Darlington, and Woo, unpublished data). 
: A. 
. 1 . 
^ 2 . 
3 . 
4. 
; 5. 
t 6. 
!( 7 . 
8 . 
LEGEND TO SOUTHERN BLOT 
NASPAH producing cells. 
N2 producing cells. 
Non- transduced hepatocytes. 
Cocultivated with NASPAH, no selection. 
Transduced with N2 , selected with G418. 
Cocultivated with N2, selected with G418. 
Transduced with NASPAH, selected with G418. 
Cocultivated with NASPAH, selected with G41 
B. LEGEND TO NORTHERN BLOT 
1. Human hepatocytes, not transduced. 
2. Murine, transduced with N2 . 
3. Human , transduced with N2 . 
4. Human, transduced with NASPAH. 
5. Mouse hepatocytes 
6. Mouse transduced with N2. 
7. Mouse transduced with NASPAH. 
i We have performed similar experiments in which human hepatocytes were transduced 
, with the N2 or NASPAH vectors and selected with G418. G418 resistant clusters of human 
i hepatocytes were obtained and proviral DNA, and RNA transcripts were identified by 
' blotting (figure 2) (Ledley, Darlington, and Woo, unpublished data). The efficiency 
I of transduction and selection of human hepatocytes was significantly worse than that 
■ observed in murine cells, possibly due to the use of post-mortem specimens and the 
: inability to use perfusion techniques for harvesting hepatocytes. 
j 
;i In summary, we believe that work from our laboratories, and others, has established 
[ the feasibility of transducing primary hepatocytes with N2 based vectors (such as LNL6) 
and achieving detectable expression of the recombinant gene from the integrated provirus 
from the LTR promoter. 
Recombinant DNA Research, Volume 14 
[793] 
