Introduction:
The current options for women fertility
preservation are scarce and some bear a great risk of reintroducing
malignancies. Developing a strategy to mature infant follicles into ovulating
follicles would eliminate this risk and provide an almost unlimited number of
potential oocytes for fertilization.
Aim:
To
elucidate the role of engineered 3D microenvironments in primordial ovarian
follicles growth and maturation
Material
and Methods:
Primordial
follicles were cultivated together with stromal cells in macroporous scaffolds
composed of pristine alginate or alginate bearing two cell adhesive peptides,
RGD and HBP. Cortical pieces of porcine ovaries were enzymatically digested and
filtered to collect follicles below 40 mm
in diameter. Cultivation proceeded 21 days. Follicle size distribution was
evaluated by a light microscope in comparison to those cultivated on
Matrigel-coated surfaces. Follicle maturation and tissue regeneration was
evaluated by a confocal microscope after fluorescently
staining with phalloidin, DAPI and GDF9.
Results:
Follicular
growth within the scaffolds demonstrated a progression towards the primary and
secondary stages to a better extent than those cultivated on Matrigel-coated
plates. In both scaffolds, the granulosa cells showed the typical cuboidal
shape, yet in the cell adhesive scaffolds, the follicles reached a maximal size
of more than 70 mm
and they had a more native ovarian tissue organization compared to those in the
pristine alginate scaffolds.
Conclusions:
This
system can be used as a first step towards the challenging task of maturing
primordial follicles to dramatically improve the chances of young oncological
women to conceive.
