Table 1.

Engraftment and transduction efficiency of SRCs

Pseudotype, incubation time, MOI, and mouse no.
Total hCD45+ cells, %
Transduced hCD45+ cells, %
Transduced hCD19+ cells, %
Transduced hCD34+ cells, %
Transduced hCD14+ cells, %
VSV-G*      
    24 h, 4       
        1A   4   11   25   40   27  
        2A   37   11.5   12   12   25  
        3A   4.4   10   14   12   10  
        4A   2   19   20   27   40  
        5A   17.5   17   13   17.5   18  
G/TPOHA*      
    24 h, 4       
        1A   18   25   13   23   23  
        2A   12   21   12   24   30  
        3A   3   36   26   33   50  
        4A   7   48   41   56   33  
        5A   5   45   43   58   51  
    72 h, 20       
        1B   21   59   58   48   63  
        2B   1   45   45   41   37  
        3B   2.5   9   7   25   44  
G/TPOHA/SCFSUx*      
    24 h, 4       
        1A   20   42   40   58   53  
        2A   17   57   40   46   42  
        3A   11   62   66   60   50  
        4A   4   69   76   83   90  
    72 h, 20       
        1B   13   67   70   48   36  
        2B   1   4   9   44   8  
G/SCFSUx*      
    24 h, 4       
        1A   1.2   38   30   36   12  
        2A   10   20   12   27   3  
        3A   1   38   21   39   36  
        4A   3.5   30   28   42   20  
    72 h, 20       
        1B   1   20   42   61   37  
        2B   1   37   51   57   41  
        3B   0.5   31   68   59   68  
G+rTPO†      
    24 h, 4       
        1A   7   22   25   36   22  
        2A   3   13   7   7.5   10  
        3A   2   14   13   27   0  
        4A   1   0   0   0   0  
G+rTPO+rSCF†      
    24 h, 4       
        1A   2   21   32   25   25  
        2A   30   35   37   42   41  
        3A   5   10   2   11   17  
        4A   21   15   8   11   20  
G+rSCF†      
    24 h, 4       
        1A   3   10   8   10   0  
        2A   24   13   11   25   11  
        3A   9.5   4   5.5   3   7.2  
        4A
 		 14
 		 3
 		 4
 		 2.5
 		 0
 
Pseudotype, incubation time, MOI, and mouse no.
Total hCD45+ cells, %
Transduced hCD45+ cells, %
Transduced hCD19+ cells, %
Transduced hCD34+ cells, %
Transduced hCD14+ cells, %
VSV-G*      
    24 h, 4       
        1A   4   11   25   40   27  
        2A   37   11.5   12   12   25  
        3A   4.4   10   14   12   10  
        4A   2   19   20   27   40  
        5A   17.5   17   13   17.5   18  
G/TPOHA*      
    24 h, 4       
        1A   18   25   13   23   23  
        2A   12   21   12   24   30  
        3A   3   36   26   33   50  
        4A   7   48   41   56   33  
        5A   5   45   43   58   51  
    72 h, 20       
        1B   21   59   58   48   63  
        2B   1   45   45   41   37  
        3B   2.5   9   7   25   44  
G/TPOHA/SCFSUx*      
    24 h, 4       
        1A   20   42   40   58   53  
        2A   17   57   40   46   42  
        3A   11   62   66   60   50  
        4A   4   69   76   83   90  
    72 h, 20       
        1B   13   67   70   48   36  
        2B   1   4   9   44   8  
G/SCFSUx*      
    24 h, 4       
        1A   1.2   38   30   36   12  
        2A   10   20   12   27   3  
        3A   1   38   21   39   36  
        4A   3.5   30   28   42   20  
    72 h, 20       
        1B   1   20   42   61   37  
        2B   1   37   51   57   41  
        3B   0.5   31   68   59   68  
G+rTPO†      
    24 h, 4       
        1A   7   22   25   36   22  
        2A   3   13   7   7.5   10  
        3A   2   14   13   27   0  
        4A   1   0   0   0   0  
G+rTPO+rSCF†      
    24 h, 4       
        1A   2   21   32   25   25  
        2A   30   35   37   42   41  
        3A   5   10   2   11   17  
        4A   21   15   8   11   20  
G+rSCF†      
    24 h, 4       
        1A   3   10   8   10   0  
        2A   24   13   11   25   11  
        3A   9.5   4   5.5   3   7.2  
        4A
 		 14
 		 3
 		 4
 		 2.5
 		 0
 

Protocol: 2E5 cord blood CD34+ cells were transduced with the indicated vector pseudotypes (G indicates VSV-G) at an MOI of 4 for 24 hours or at an MOI of 20 for 72 hours. After 72 hours or 24 hours of incubation with vector, CD34+ cells were washed and injected by tail vain injection into sublethally irradiated NOD/SCID mice. The animals were humanely killed 7 weeks later and the BM was harvested and assessed for levels of human engraftment and GFP-expressing human cells. Multilineage engraftment was demonstrated by lineage-positive markers as indicated and for each lineage the percentage EGFP+ cells was analyzed by FACS. Independent experiments were performed with different CD34+ CB samples and different preparations of each vector.

*

Incubation with cytokine displaying lentivectors without addition of recombinant cytokines.

†

Incubation with lentivectors and addition of indicated cytokines (TPO, 10 ng/mL; SCF, 50 ng/mL; Flk-3, 100 ng/mL).

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