Cs. Wright et al., Effects of follicle-stimulating hormone and serum substitution on the in-vitro growth of human ovarian follicles, HUM REPR, 14(6), 1999, pp. 1555-1562
In-vitro maturation (IVM) of human ovarian follicles and oocytes could bene
fit infertile women, and allow the development of in-vitro systems for the
study of human follicular development. Little is known about the initiation
of growth of primordial follicles and the regulation of early folliculogen
esis. An ovarian tissue-slice culture system was used to examine the effect
s of media composition, follicle stimulating hormone (FSH) and serum substi
tution on the development of small human follicles in vitro. Human ovarian
cortex biopsies were cut into small pieces and cultured for 5, 10 or 15 day
s. Control (non-cultured) and cultured tissue was fixed, serially sectioned
, and stained. The follicles contained within the tissue pieces were counte
d, measured, and assessed for stage of development and viability. Compariso
n of the ability of alpha-minimum essential medium (alpha-MEM), Waymouth's,
or Earle's balanced salt solution (EBSS) culture media (all with 10% human
serum) to support follicle growth demonstrated significantly increased ini
tiation and growth of follicles in alpha-MEM during the first 10 days of cu
lture. The supplementation of alpha-MEM with 300 mIU/ml FSH significantly r
educed levels of atresia and increased the mean diameter of healthy follicl
es. Follicles in tissue cultured for 10 days with human serum albumin and I
TS (insulin/transferrin/selenium mix) were significantly larger, more devel
oped and showed significantly less atresia than those cultured with serum a
lone. Primordial to small preantral follicles can be grown under serum-subs
tituted conditions in tissue-slice culture, and are responsive to FSH, whic
h is thought to be acting mainly as a survival factor at these early stages
.