Title | Activated ovarian endothelial cells promote early follicular development and survival. |
Publication Type | Journal Article |
Year of Publication | 2017 |
Authors | Kedem A, Aelion-Brauer A, Guo P, Wen D, Ding B-S, Lis R, Cheng D, Sandler VM, Rafii S, Rosenwaks Z |
Journal | J Ovarian Res |
Volume | 10 |
Issue | 1 |
Pagination | 64 |
Date Published | 2017 Sep 19 |
ISSN | 1757-2215 |
Keywords | Animals, Apoptosis, Cell Communication, Cell Culture Techniques, Cell Proliferation, Cells, Cultured, Endothelial Cells, Estradiol, Female, Fibroblasts, Granulosa Cells, Mice, Oocytes, Ovarian Follicle |
Abstract | <p><b>BACKGROUND: </b>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.</p><p><b>METHODS: </b>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.</p><p><b>RESULTS: </b>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.</p><p><b>CONCLUSIONS: </b>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.</p><p><b>CAPSULE: </b>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.</p> |
DOI | 10.1186/s13048-017-0354-z |
Alternate Journal | J Ovarian Res |
PubMed ID | 28927438 |
PubMed Central ID | PMC5605973 |