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Table 3.

Hedgehog signalling affects HSC activity in AGM explants

Embryonic dayTissueFactor or antibodyConcentration (ng/ml)Number repopulated/number transplanted% Recipients repopulated% Chimerism
Early E10 AGM Ihh 0/9 
   20 4/10 40 100, 89, 99, 91 
   200 0/12 
Early E10 AGM Shh 0/3 
   20 4/10 40 95, 89, 95, 33 
   200 0/8 
Early E10 AGM:gut IgG 50 2/8 25 60, 99 
  Anti-Hh 50 0/7 
Late E10 AGM:gut IgG 50 3/5 60 11, 47, 83 
  Anti-Hh 50 1/6 17 99 
E11 AGM IgG 50 4/11 36 >10, >10, >10, >10 
  Anti-Hh 50 3/12 25 >10, >10, >10 
Embryonic dayTissueFactor or antibodyConcentration (ng/ml)Number repopulated/number transplanted% Recipients repopulated% Chimerism
Early E10 AGM Ihh 0/9 
   20 4/10 40 100, 89, 99, 91 
   200 0/12 
Early E10 AGM Shh 0/3 
   20 4/10 40 95, 89, 95, 33 
   200 0/8 
Early E10 AGM:gut IgG 50 2/8 25 60, 99 
  Anti-Hh 50 0/7 
Late E10 AGM:gut IgG 50 3/5 60 11, 47, 83 
  Anti-Hh 50 1/6 17 99 
E11 AGM IgG 50 4/11 36 >10, >10, >10, >10 
  Anti-Hh 50 3/12 25 >10, >10, >10 

E10 AGM explants (Ln72) were cultured for 3 days and cells transplanted into irradiated adult recipients (B10 × CBAF1). Recipients received 2-2.5 embryo equivalents of AGM cells. At >4 months after transplantation, peripheral blood DNA PCR was performed for the Ln72transgene. Percentage donor contribution was determined by comparison of the specific signal that was normalized for DNA content (myogenincontrol) with serial dilutions (100, 60, 30, 10, 6, 1 and 0%) of control transgenic DNA. Total number of experiments=14. For each condition n=2-4.

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