pore size, Millipore Corp, Billerica, MA, US) at 37˚C and 5% CO2 for 24 hours
in the presence (n=57) and absence (n=57) of LPA. The culture media consisted of 300 µl
α-MEM medium supplemented with 5% foetal bovine serum; 1% insulin, transferrin, and
selenium (Invitrogen, UK), and 100 mIU/ml recombinant follicle stimulating hormone
(Serono, Switzerland). The treated group had 20 µM LPA (INstruchemie, The Netherlands)
added to the culture medium (31 (link)).
The cultured ovaries were observed under an inverted
microscope and some of the cultured ovaries were
considered for assessment of ovarian cell survival using
Calcein AM (n=6 for the LPA-treated group and n=6 for
the non-treated group) and for morphological analysis
with haematoxyline and eosin (n=5 for the LPA-treated
group and n=5 for the non-treated group). The other
cultured ovaries were encapsulated with sodium alginate
and then transplanted under kidney capsule (n=45 for the
LPA-treated group and n=45 for the non-treated group).