Human Reproduction, Vol 12, 1993-2001, Copyright © 1997 by Oxford University Press
SA Wandji, V Srsen, PW Nathanielsz, JJ Eppig and JE Fortune
Factors that cause some primordial follicles to enter the growth phase
while the others remain quiescent are unknown. The hypothesis was tested
that primate primordial follicles can survive and initiate growth in vitro
in serum-free medium. Superficial pieces of ovarian cortex, containing
mostly primordial follicles, were obtained from baboon fetuses during late
gestation and cultured for 0, 2, 4, 7, 10 or 20 days in Waymouth MB 752/1
medium supplemented with insulin, transferrin, selenium, linoleic acid, and
bovine serum albumin (ITS +). Histological examination of cortical pieces
revealed that after 2 and 4 days in culture, the total number of primordial
follicles had decreased by 55 and 76% (P < 0.01) respectively, relative
to day 0 of culture. This was associated with a sustained, 5- to 8-fold
increase in total primary follicles (P < 0.01) beginning on day 2 of
culture. There was also a gradual increase in the total number of early
secondary and secondary follicles. The average diameter of follicles and
oocytes increased gradually throughout culture for all follicular
categories (P < 0.01), except secondary follicles and oocytes.
Immunohistochemical localization of proliferating cell nuclear antigen
(PCNA), a marker for cell proliferation and growth, showed that PCNA was
generally absent in primordial follicles on day 0, but was observed after 2
or 4 days in culture in both granulosa cells and oocytes of most growing
follicles. Comparison of cortical pieces cultured for 10 or 20 days with
ITS + versus 10% fetal bovine serum (FBS) showed a more pronounced decrease
in the numbers of primordial follicles and more primary, early secondary
and secondary follicles in ITS + compared to FBS-treated cortical pieces (P
< 0.01 at 20 days). These results show that primordial follicles from
non-human primates can survive and develop to the secondary stage in vitro
in serum-free conditions.
ARTICLES
Initiation of growth of baboon primordial follicles in vitro
Department and Section of Physiology, College of Veterinary Medicine, Cornell University, Ithaca, New York 14853, USA.
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