Table 6.

Effects of GSK3 inhibition and p53 deficiency on EG cell derivation from E11.5 PGCs

A
Growth factors Treatment Seeded PGC (embryo/well) EG cell-positive wellsχ 2 test DMSO
bFGF, SCF, LIFDMSO0.025/12-
meBIO0.025/12nsd
BIO0.025/12nsd
SCF, LIFDMSO0.10/12-
meBIO0.10/12nsd
BIO 0.1 0/12 nsd
B
Genotype Growth factors Seeded PGC (embryo/well) EG cell-positive wellsχ 2 test −/−
+/+ (n=2)bFGF, SCF, LIF0.021/12<0.005
+/− (n=11)0.029/66<0.005
−/− (n=4)0.0222/24
+/+ (n=5)SCF, LIF0.10/25<0.01
+/− (n=12)0.10/60<0.005
−/− (n=7)0.18/35
  • (A) E11.5 wild-type PGCs were cultured with the GSK3 inhibitor BIO, its inactivated derivative Me-BIO (methyl-BIO) or DMSO during primary culture. EG cell derivation was carried out as described in Fig. 2A.

    (B) p53 heterozygous females were crossed with heterozygous males. Gonads were isolated at E11.5. Cell suspensions from the individual embryos were seeded onto Sl/Sl4-m220 feeder cells and cultured with growth factors as indicated.