In vitro maturation of oocytes via the pre-fabricated self-assembled artificial human ovary

Krotz SP, Robins JC, Ferruccio TM, Moore R, Steinhoff MM, Morgan JR, Carson S.
Source
Division of Reproductive Endocrinology & Infertility, Department of Obstetrics & Gynecology, Women & Infants’ Hospital, Alpert Medical School, Brown University, Providence, RI 02905, USA.

Abstract
PURPOSE:
create a 3-Dimensional artificial human ovary to mature human oocytes.

METHODS:
theca and granulosa cells were isolated from antral follicles of reproductive-aged women, seeded into micro-molded gels and self-assembled into complex 3D microtissues. Immunohistochemistry and live-dead staining confirmed theca cell identity and cellular viability at one week respectively. Placement of granulosa cell spheroids or cumulus-oocyte complexes into theca cell honeycomb openings resulted in creation of an artificial human ovary. Oocytes from this construct were assessed for polar body extrusion.

RESULTS:
theca and granulosa cells self-assembled into complex microtissues, remaining viable for one week. At 72 h after artificial human ovary construction, theca cells completely surrounded the granulosa spheroids or COCs without stromal invasion or disruption. Polar body extrusion occurred in one of three COCs assessed.

CONCLUSIONS:
an artifical human ovary can be created with self-assembled human theca and granulosa cell microtissues, and used for IVM and future oocyte toxicology studies.

3D Petri Dishes™ used in this paper (please click catalog numbers for detailed product descriptions):
Catalog # 12-256, and 24-96 are the MicroTissues, Inc products that grow small spheroids. Catalog # 12-81 and 24-35 are used to grow larger spheroids. Catalog # 24-H is used to form a small multi-cellular honeycomb structure.

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