We differentiated hESCs to PE following established methods
[13]. The PE was then transplanted into db/+ female mice
(db/+PE), whereas db/+ mice receiving no transplants (db/+)
and wild-type (wt) mice were used as controls. Four weeks
after transplant, breeding was conducted overnight in a 1:2
ratio; mating was confirmed by the presence of a vaginal
mucous plug the following morning, which represented
gestation day (GD) 0 (GD0). Blood glucose levels, plasma
insulin levels, and body weight were measured at the
following stages: prior to PE transplant (day 0), 4 weeks after
transplant (day 28), and at GD0, GD10, and GD20. Numbers of
offsprings were counted at birth for each experimental group
of dams. Body weight of offsprings was measured at days 0, 7,
and 14 after birth.