, Volume 27, Issue 12, pp 743-750
Date: 25 Aug 2010

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

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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.

Capsule

An artificial human ovary can be created using self-assembled 3-dimensional theca & granulosa cell constructs to mature human oocytes in vitro