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J Ovarian Res. 2017 Sep 19;10(1):64. doi: 10.1186/s13048-017-0354-z.

Activated ovarian endothelial cells promote early follicular development and survival.

Author information

1
The Center for Reproductive Medicine and Infertility, Weill Cornell Medical College, New York, NY, 10021, USA. kedem2001@gmail.com.
2
Ansary Stem Cell Center for Regenerative Medicine at Weill Medical College of Cornell University, New York, USA. kedem2001@gmail.com.
3
The Center for Reproductive Medicine and Infertility, Weill Cornell Medical College, New York, NY, 10021, USA. aaelion@gmail.com.
4
Ansary Stem Cell Center for Regenerative Medicine at Weill Medical College of Cornell University, New York, USA.
5
The Center for Reproductive Medicine and Infertility, Weill Cornell Medical College, New York, NY, 10021, USA.

Abstract

BACKGROUND:

New data suggests that endothelial cells (ECs) elaborate essential "angiocrine factors". The aim of this study is to investigate the role of activated ovarian endothelial cells in early in-vitro follicular development.

METHODS:

Mouse ovarian ECs were isolated using magnetic cell sorting or by FACS and cultured in serum free media. After a constitutive activation of the Akt pathway was initiated, early follicles (50-150 um) were mechanically isolated from 8-day-old mice and co-cultured with these activated ovarian endothelial cells (AOEC) (n = 32), gel (n = 24) or within matrigel (n = 27) in serum free media for 14 days. Follicular growth, survival and function were assessed.

RESULTS:

After 6 passages, flow cytometry showed 93% of cells grown in serum-free culture were VE-cadherin positive, CD-31 positive and CD 45 negative, matching the known EC profile. Beginning on day 4 of culture, we observed significantly higher follicular and oocyte growth rates in follicles co-cultured with AOECs compared with follicles on gel or matrigel. After 14 days of culture, 73% of primary follicles and 83% of secondary follicles co-cultured with AOEC survived, whereas the majority of follicles cultured on gel or matrigel underwent atresia.

CONCLUSIONS:

This is the first report of successful isolation and culture of ovarian ECs. We suggest that co-culture with activated ovarian ECs promotes early follicular development and survival. This model is a novel platform for the in vitro maturation of early follicles and for the future exploration of endothelial-follicular communication.

CAPSULE:

In vitro development of early follicles necessitates a complex interplay of growth factors and signals required for development. Endothelial cells (ECs) may elaborate essential "angiocrine factors" involved in organ regeneration. We demonstrate that co-culture with ovarian ECs enables culture of primary and early secondary mouse ovarian follicles.

KEYWORDS:

Endothelial cells; Follicle culture; Follicular activation; in-vitro maturation

PMID:
28927438
PMCID:
PMC5605973
DOI:
10.1186/s13048-017-0354-z
[Indexed for MEDLINE]
Free PMC Article

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